CN105999295A - Tolerogenic synthetic nanocarriers to reduce immune responses to therapeutic proteins - Google Patents

Tolerogenic synthetic nanocarriers to reduce immune responses to therapeutic proteins Download PDF

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Publication number
CN105999295A
CN105999295A CN201610345030.5A CN201610345030A CN105999295A CN 105999295 A CN105999295 A CN 105999295A CN 201610345030 A CN201610345030 A CN 201610345030A CN 105999295 A CN105999295 A CN 105999295A
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China
Prior art keywords
synthetic nanocarriers
population
cells
antigen
subject
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CN201610345030.5A
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Chinese (zh)
Inventor
克里斯托弗·弗雷泽
岸本·隆·慧
罗伯托·A·马尔多纳多
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西莱克塔生物科技公司
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Priority to US201161480946P priority Critical
Priority to US201161513514P priority
Priority to US201161531215P priority
Priority to US201161531147P priority
Priority to US201161531209P priority
Priority to US201161531194P priority
Priority to US201161531153P priority
Priority to US201161531204P priority
Priority to US201161531175P priority
Priority to US201161531168P priority
Priority to US201161531180P priority
Priority to US201161531164P priority
Application filed by 西莱克塔生物科技公司 filed Critical 西莱克塔生物科技公司
Priority to CN201280020311.3A priority patent/CN103501820B/en
Publication of CN105999295A publication Critical patent/CN105999295A/en

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Abstract

The application relates to tolerogenic synthetic nanocarriers to reduce immune responses to therapeutic proteins, and discloses methods of synthesizing nanocarriers and related compositions, wherein the methods include providing an immunosuppressor that can produce tolerogenic immune responses (e,g. antigen-specific effector T cell deficiency), and MHC I-type and/or MHC II-type restricted epitopes.

Description

用于效应性τ细胞的抗原特异性缺失的致耐受性合成纳米载体 Induced by specific deletion of synthetic nanocarriers for effector cell antigen tolerance τ

[0001] 本申请是申请日为2012年4月27日、申请号为"201280020311.3"、发明名称为"用于效应性Τ细胞的抗原特异性缺失的致耐受性合成纳米载体"的中国专利申请的分案申请, 原申请是国际申请PCT/US2012/035555的中国国家阶段申请。 Chinese Patent [0001] This application is filed April 27, 2012, application number "201280020311.3", entitled "tolerance induced by synthetic nanocarriers for specific deletion of antigen Τ effector cells" in divisional application filed, the original application is the Chinese national phase application of international application PCT / US2012 / 035555 of.

[000^ 相关申请 [000 ^ RELATED APPLICATIONS

[0003] 本申请根据美国法典第35篇第119条要求2011年4月29日提交的美国临时申请61/ 480,946、2011年7月29日提交的美国临时申请61/513,514、2011年9月6日提交的美国临时申请61/531,147、2011年9月6日提交的美国临时申请61/531,153、2011年9月6日提交的美国临时申请61/531,164、2011年9月6日提交的美国临时申请61/531,168、2011年9月6日提交的美国临时申请61/531,175、2011年9月6日提交的美国临时申请61/531,180、2011年9月6日提交的美国临时申请61/531,194、2011年9月6日提交的美国临时申请61/531,204、2011 年9月6日提交的美国临时申请61/531,209、2011年9月6日提交的美国临时申请61/531,215 的权益,运些临时申请各自的全部内容通过引用结合在此。 [0003] This application claims priority under US Code Title 35 Section 119 of the United States April 29, 2011 Provisional Application 61 / 480,946, United States July 29, 2011 Provisional Application 61/513, 514, 2011, September 6 US provisional application filed 61 / 531,147, US provisional application No. 61 September 6, 2011 submitted / 531,153, US provisional application No. 61 September 6, 2011 submitted / 531,164, September 2011 6 US provisional application 61 / 531,168, United States September 6, 2011 provisional application 61 / 531,175, United States September 6, 2011 provisional application 61 / 531,180, September 2011 filed May 6, US provisional application No. 61 / 531,194, United States September 6, 2011 filed provisional application 61 / 531,204,2011 on September 6 US provisional application No. 61 / 531,209, 2011 US September 6 provisional application 61 / 531,215, and shipped some provisional application entire contents of each incorporated herein by reference.

技术领域 FIELD

[0004] 本发明设及给予具有免疫抑制剂和抗原的MHC I类限制性表位和/或MHC II类限制性表位的合成纳米载体组合物的方法、W及有关组合物,当分别WMHC I类分子或血1C II 类分子呈递时运些表位是抗原特异性效应性Τ细胞的同源表位。 [0004] The present invention is provided and administered MHC class I restricted epitope and / or MHC II-restricted epitopes and antigens having immunosuppressant synthetic nanocarriers compositions methods, W, and related compositions, respectively, when WMHC class I molecules or blood 1C II molecules presenting these epitopes fortune antigen-specific effector cells Τ cognate epitope. 在一些实施例中,运些方法允许通过APC有效摄取来使免疫应答向有利于缺失抗原特异性效应性Τ细胞的方向转变。 In some embodiments, these methods allow the operation to make efficient uptake by the APC immune response in favor of the deletion of antigen-specific effector cells Τ transition.

背景技术 Background technique

[0005] 用于产生与所不希望的免疫应答相关联的免疫抑制的常规策略是基于广泛作用的免疫抑制性药物。 Conventional immunization strategies [0005] for generating undesired immune response associated with the inhibition of immunosuppressive drugs it is based on extensive action. 另外,为了维持免疫抑制,免疫抑制剂药物疗法通常是一个终生的命题。 Further, in order to maintain immunosuppression, immunosuppressant drug therapy usually is a lifetime proposition. 令人遗憾的是,使用广泛作用的免疫抑制剂与严重副作用(如肿瘤、感染、肾毒性W及代谢素乱)的风险相关联。 Unfortunately, the widespread use of immunosuppressive effects and serious side effects (such as cancer, infection, renal toxicity W and metabolic hormone disorder) risk associated. 因此,新的免疫抑制剂疗法将是有益的。 Therefore, the new immunosuppressant therapy will be beneficial.

发明内容 SUMMARY

[0006] 在一个方面,提供了一种包括根据一个治疗方案向受试者给予一种组合物的方法,该治疗方法先前已经显示在一个或多个测试受试者中降低抗原特异性效应性Τ细胞的数目或活性;其中该组合物包含:(i)第一群体的合成纳米载体和(ii)第二群体的合成纳米载体,运些第一群体的合成纳米载体与免疫抑制剂偶联,运些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位或MHC II类限制性表位偶联。 [0006] In one aspect, there is provided a composition comprising administering a therapeutic composition in accordance with one embodiment of the subject method, the method of treatment has previously been shown to reduce antigen-specific effects in one or more subject test the number or activity of Τ cells; wherein the composition comprises: synthesis nanocarrier (i) a first population of the synthetic nanocarriers and (ii) a second population, coupled with the synthetic nanocarriers transport immunosuppressant plurality of first groups , some synthetic nanocarriers shipped second population with an MHC class I-restricted epitopes of an antigen or MHC class II restricted epitopes conjugated. 在另一个方面,提供了一种包括通过向一个或多个测试受试者给予一种组合物降低在一个或多个测试受试者中的抗原特异性效应性T细胞的数目或活性的方法;该组合物包含:(i)第一群体的合成纳米载体和(ii)第二群体的合成纳米载体,运些第一群体的合成纳米载体与免疫抑制剂偶联,运些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位或MHC II类限制性表位偶联。 In another aspect, there is provided a method comprising the number or activity of effector antigen-specific T cells in one or more test subject by administering a composition to a subject to reduce one or more test ; the composition comprising: synthesis nanocarrier (i) a first population of the synthetic nanocarriers and (ii) a second population, coupled with the synthetic nanocarriers transport immunosuppressant plurality of first groups of the second population of these transport class restricted epitopes conjugated synthetic nanocarriers MHC class I restricted epitopes of an antigen or MHC II. 在另一个方面,提供了一种包括向受试者给予一种组合物的方法,该组合物包含:(i)第一群体的合成纳米载体和(ii)第二群体的合成纳米载体,运些第一群体的合成纳米载体与免疫抑制剂偶联,运些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II 类限制性表位偶联。 In another aspect, there is provided a composition comprising administering to a subject methods, the composition comprising: Synthesis nanocarrier (i) a first population of the synthetic nanocarriers and (ii) a second population, transport these synthetic nanocarriers coupled with a first group of immunosuppressants, these synthetic nanocarriers shipped second population with an MHC class I-restricted epitopes of an antigen and / or MHC class II restricted epitopes conjugated.

[0007] 优选地,当运些表位分别肥I类或MHC II类分子呈递时,它们是抗原特异性效应性T细胞的同源表位。 [0007] Preferably, when these epitopes are transported fertilizer class I or class II MHC molecules presenting their cognate epitopes are antigen-specific effector T cells. 在一个实施例中,运些效应性T细胞是CD4+或CD8巧细胞。 In one embodiment, the transport of these effector T cells are CD4 + or CD8 cells clever.

[0008] 在一个实施例中,该第一群体和该第二群体是相同的群体。 [0008] In one embodiment, the first population and the second population are identical groups. 在另一个实施例中,该第一群体和该第二群体是不同的群体。 In another embodiment, the first groups and the second groups are different groups.

[0009] 在另一个实施例中,该方法进一步包括提供或鉴定该受试者。 [0009] In another embodiment, the method further comprises providing or identifying the subject.

[0010] 在另一个实施例中,该抗原是一种治疗性蛋白、一种自身抗原或一种过敏原,或与炎性疾病、自身免疫性疾病、器官或组织排斥或移植物抗宿主病相关联。 [0010] In another embodiment, the therapeutic protein is an antigen, a self antigen or an allergen, or an inflammatory disease, an autoimmune disease, organ or tissue rejection or graft versus host disease Associated.

[0011] 在另一个实施例中,该方法进一步包括在给予该组合物之前和/或之后评定该受试者中的抗原特异性效应性T细胞的数目或活性。 [0011] In another embodiment, the method further includes assessing the number or activity of the subject antigen-specific effector T cells and / or before and after administration of the composition.

[0012] 在另一个实施例中,该受试者具有或正处于具有一种炎性疾病、一种自身免疫性疾病、一种过敏症、器官或组织排斥或移植物抗宿主病的风险。 [0012] In another embodiment, the subject has or is in having an inflammatory disease, an autoimmune disease, an allergic disease, organ or tissue rejection or graft versus host disease risk. 在另一个实施例中,该受试者已经经历或将要经历移植。 In another embodiment, the subject has undergone or will undergo transplantation. 在另一个实施例中,该受试者具有或正处于具有针对一种治疗性蛋白的一种所不希望的免疫应答的风险,该治疗性蛋白是正被给予或将要被给予该受试者的。 In another embodiment, the subject has or is at risk of having a therapeutic protein against one of the undesirable immune response, the therapeutic protein is being administered or will be administered to the subject .

[0013] 在另一个实施例中,该方法进一步包括给予一种可移植的移植物或治疗性蛋白。 [0013] In another embodiment, the method further comprises administering a therapeutic graft, or a portable protein. 在另一个实施例中,向该受试者给予一个或多个维持剂量的该组合物,该组合物包含运些第一群体和第二群体的合成纳米载体。 In another embodiment, administering to the subject one or more maintenance dose of the composition, the composition comprising a plurality of first groups and the second operation group of synthetic nanocarriers.

[0014] 在另一个实施例中,该给予是通过静脉内、腹膜内、透粘膜、经口、皮下、肺部、鼻内、皮内或肌肉内给予而进行的。 [0014] In another embodiment, the administration is by intravenous, intraperitoneal, transmucosal, oral, subcutaneous, pulmonary, intranasal, intradermal administration or intramuscular carried out. 在另一个实施例中,该给予是通过吸入或静脉内、皮下或透粘膜给予而进行的。 In another embodiment, the administration is by inhalation or intravenous, subcutaneous or transmucosal administration carried out. 在另一个实施例中,当该治疗性蛋白作为一个或多个单元被提供时, 该可移植的移植物或治疗性蛋白的给予是通过肠胃外、动脉内、鼻内或静脉内给予或通过注射至淋己结或眼前房或通过局部给予至感兴趣的器官或组织而进行的。 In another embodiment, when the therapeutic protein is provided as one or more units, the graft or implantable therapeutic protein is administered by parenteral, intraarterial, intranasal or intravenous administration or by injection into the lymph nodes or hexyl anterior chamber or by topical administration to an organ or tissue of interest is carried out.

[0015] 在另一个实施例中,运些免疫抑制剂包括一种抑制素、一种mTOR抑制剂、一种TGF- 0信号剂、一种皮质类固醇、一种线粒体功能抑制剂、一种P38抑制剂、一种NF-κβ抑制剂、一种腺巧受体激动剂、一种前列腺素E2激动剂、一种憐酸二醋酶4抑制剂、一种HDA巧Φ制剂或一种蛋白酶体抑制剂。 [0015] In another embodiment, these immunosuppressive agents include one transport statin, one mTOR inhibitor, a signal reagent TGF- 0, one corticosteroid, a mitochondrial function inhibitor, a P38 inhibitor, a NF-κβ inhibitor, a clever adenovirus receptor agonist, a prostaglandin E2 agonist A pity acid esterase 4 inhibitor, a clever HDA or one formulation proteasome Φ inhibitors. 在另一个实施例中,该mTOR抑制剂是雷帕霉素或一种雷帕霉素类似物。 In another embodiment, the mTOR inhibitor is rapamycin or one rapamycin analog.

[0016] 在另一个实施例中,在第一和/或第二群体的合成纳米载体上的该免疫抑制剂和/ 或表位、或包含运些表位的蛋白质、多肤或肤的平均负载量是在0.0001%与50% (重量/重量)之间。 [0016] In embodiments, the immunosuppressive agent on synthetic nanocarriers of the first and / or second groups and / or epitope in another embodiment, or more epitope comprising transport protein, on average skin or the skin loading is between 0.0001% and 50% (wt / wt). 在另一个实施例中,在第一和/或第二群体的合成纳米载体上的该免疫抑制剂和/ 或表位、或包含运些表位的蛋白质、多肤或肤的平均负载量是在0.1%与10% (重量/重量) 之间。 Embodiment, the immunosuppressive agent on synthetic nanocarriers of the first and / or second groups and / or epitope in another embodiment, a protein or an epitope comprising some transport, average load multiple skin or skin is between 0.1% and 10% (wt / wt).

[0017] 在另一个实施例中,该第一群体和/或第二群体的合成纳米载体包括脂质纳米颗粒、聚合物纳米颗粒、金属纳米颗粒、基于表面活性剂的乳液、树枝状化合物、己基球、纳米线、病毒样颗粒或肤或蛋白质颗粒。 [0017] In another embodiment, the first population and / or synthetic nanocarriers second population comprises lipid nanoparticles, polymeric nanoparticles, metal nanoparticles, emulsion based surfactants, dendrimers, hexyl ball, nanowires, skin, or virus-like particle or protein particles. 在另一个实施例中,该第一群体和/或第二群体的合成纳米载体包括脂质纳米颗粒。 In another embodiment, the first population and / or synthetic nanocarriers second population comprises lipid nanoparticles. 在另一个实施例中,该第一群体和/或第二群体的合成纳米载体包括脂质体。 In another embodiment, the first population and / or second population of synthetic nanocarriers include liposomes. 在另一个实施例中,该第一群体和/或第二群体的合成纳米载体包括金属纳米颗粒。 In another embodiment, the first population and / or second population of synthetic nanocarriers include metal nanoparticles. 在另一个实施例中,运些金属纳米颗粒包括金纳米颗粒。 In another embodiment, the transport of these metals nanoparticles comprise gold nanoparticles. 在另一个实施例中,该第一群体和/或第二群体的合成纳米载体包括聚合物纳米颗粒。 In another embodiment, the first population and / or synthetic nanocarriers second population comprises polymeric nanoparticles. 在另一个实施例中,该聚合物纳米颗粒包括聚合物,该聚合物是一种非甲氧基封端的普朗尼克聚合物。 In another embodiment, the polymer nanoparticles comprise a polymer, the polymer is a non-methoxy-terminated Pluronic polymers. 在另一个实施例中,运些聚合物纳米颗粒包括聚醋、与聚酸偶联的聚醋、聚氨基酸、聚碳酸醋、聚缩醒、聚缩酬、多糖、聚乙基啜挫嘟、或聚乙締亚胺。 In another embodiment, these operation include polyester polymeric nanoparticles, coupled with the polyacid polyester, polyamino acids, polycarbonates, polycondensation wake up, pay polycondensation, polysaccharides, polyethyl sip beep setback, association or polyethylene imine. 在另一个实施例中,聚醋包括聚(乳酸)、聚(乙醇酸)、聚(乳酸-乙醇酸)共聚物或聚己酸内醋。 In another embodiment, the polyester comprises a poly (lactic acid), poly (glycolic acid), poly - inner (lactic-glycolic acid) or polycaprolactone vinegar. 在另一个实施例中,运些聚合物纳米颗粒包括聚醋W及与聚酸偶联的聚醋。 In another embodiment, these polymer nanoparticles comprising transport polyester and polyester W and coupled to a polyacid. 在另一个实施例中,该聚酸包括聚乙二醇或聚丙二醇。 In another embodiment, the polyacid comprises polyethylene glycol or polypropylene glycol.

[0018] 在另一个实施例中,使用对该第一和/或第二群体的合成纳米载体进行的动态光散射而获得的粒度分布的平均值是大于lOOnm的直径。 [0018] In another embodiment, the dynamic light scattering using a first and / or second population of synthetic nanocarriers performed and the average particle size distribution is obtained of a diameter greater than lOOnm. 在另一个实施例中,该直径大于150nm。 In another embodiment, the diameter is greater than 150nm. 在另一个实施例中,该直径大于200nm。 In another embodiment, the diameter is greater than 200nm. 在另一个实施例中,该直径大于250nm。 In another embodiment, the diameter is greater than 250nm. 在另一个实施例中,该直径大于300nm。 In another embodiment, the diameter is greater than 300nm.

[0019] 在另一个实施例中,该第一群体和/或第二群体的合成纳米载体的长宽比大于1: 1、1:1.2、1:1.5、1:2、1:3、1:5、1:7或1:10。 [0019] In another embodiment, the aspect ratio of the first population and / or synthetic nanocarriers second population is greater than 1: 1, 1: 1.2, 1: 1.5, 1: 2,1: 3,1 : 5, 1: 7 or 1:10.

[0020] 在另一个方面,提供了一种方法,该方法包括:(i)产生第一群体的合成纳米载体和(ii)产生第二群体的合成纳米载体,运些第一群体的合成纳米载体与免疫抑制剂偶联, 运些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位或MHC II类限制性表位偶联;并且评估运些第一群体和第二群体的合成纳米载体对抗原特异性效应性T细胞的数目或活性的作用。 [0020] In another aspect, there is provided a method comprising: (i) generating a first synthetic nanocarriers groups and (ii) generating a second synthetic nanocarriers groups, a first group of these transport Nanometer immunosuppressant carrier coupled operation of these synthetic nanocarriers second population with an MHC class I-restricted epitopes of an antigen or MHC class II restricted epitopes conjugated; and evaluating operators of these first and second groups effect on the number or activity of the antigen-specific effector T cells synthetic nanocarriers groups. 在一个实施例中,运种作用在将运些第一群体和第二群体的合成纳米载体给予该受试者之后进行评估。 After assessed in one embodiment, the transport operation some kind of a role in the first population and the second population of synthetic nanocarriers administered to the subject. 在另一个实施例中,使用获自该受试者的一个样品进行评估。 In another embodiment, using a sample obtained from the assessment of the subject.

[0021] 在另一个实施例中,该第一群体和第二群体是相同的群体。 [0021] In another embodiment, the first population and the second population are identical groups. 在另一个实施例中,该第一群体和第二群体是不同的群体。 In another embodiment, the first population and the second population are different groups.

[0022] 在另一个实施例中,该方法进一步包括制造一种包含运些第一群体和第二群体的合成纳米载体的剂型。 [0022] In another embodiment, the method further comprises producing a dosage form comprising a plurality of first groups and the second operation group of synthetic nanocarriers. 在另一个实施例中,该方法进一步包括制造一种包含运些第一群体和第二群体的合成纳米载体的组合物、或可供受试者给予的剂型。 In another embodiment, the method further comprises producing a composition comprising a plurality of first groups and the second operation group of synthetic nanocarriers, subject, or a dosage form for administration. 在另一个实施例中,该方法进一步包括在给予该组合物或剂型之前和/或之后评定该受试者中的抗原特异性效应性T细胞的数目或活性。 In another embodiment, the method further includes assessing the number or activity of the subject antigen-specific effector T cells and / or before and after administration of the composition or dosage form.

[0023] 优选地,当运些表位分别WifflC I类或MHC II类分子呈递时,它们是抗原特异性效应性T细胞的同源表位。 [0023] Preferably, when these epitopes are transported WifflC class I or MHC II molecules presentation, they are homologous epitope-specific effector T cells. 在一个实施例中,运些效应性T细胞是CD4+或CD8巧细胞。 In one embodiment, the transport of these effector T cells are CD4 + or CD8 cells clever.

[0024] 在另一个实施例中,产生的运些第一群体和第二群体的合成纳米载体是如在此提供的任何运些方法和组合物中所定义的。 [0024] In another embodiment, the operation of these synthetic nanocarriers first population and second population produced is transported to any of these methods and compositions provided herein defined.

[0025] 在另一个方面,提供了一种生产一种组合物或剂型的方法,该方法包括:(i)将第一群体的合成纳米载体与免疫抑制剂偶联;(ii)将第二群体的合成纳米载体与一种抗原的Μ肥I类限制性表位和/或MHC II类限制性表位偶联;并且(iii)评估运些第一群体和第二群体的合成纳米载体对抗原特异性效应性T细胞的数目或活性的作用。 [0025] In another aspect, there is provided a process for producing a composition or dosage form, the method comprising: (i) Coupling of the synthetic nanocarriers immunosuppressant first population; (ii) a second synthetic nanocarriers groups and Μ fertilizer class I restricted epitopes of an antigen and / or MHC class II restricted epitopes coupling; and (iii) evaluating operators of these synthetic nanocarriers first population and second population of role of antigen-specific number or activity of effector T cells. 在一个实施例中,运种作用在将运些第一群体和第二群体的合成纳米载体给予该受试者之后进行评估。 After assessed in one embodiment, the transport operation some kind of a role in the first population and the second population of synthetic nanocarriers administered to the subject. 在另一个实施例中,使用获得自该受试者的一个样品进行评估。 In another embodiment, using the obtained evaluated from a sample of the subject. 在另一个方面,该工艺包括在此提供的任何方法或工艺中所定义的步骤。 In another aspect, the process comprising the steps of any method or process provided herein defined.

[0026] 在另一方面,提供了一种通过在此提供的任何方法或工艺可获得的组合物或剂型。 [0026] In another aspect, there is provided a composition or a dosage form by any method or process provided herein obtainable.

[0027] 在另一个方面,该组合物包含:第一群体的合成纳米载体;第二群体的合成纳米载体;W及一种药学上可接受的赋形剂,运些第一群体的合成纳米载体与免疫抑制剂偶联,运些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联。 [0027] In another aspect, the composition comprising: a first synthetic nanocarrier groups; second population of synthetic nanocarriers; Nanometer pharmaceutically W and a pharmaceutically acceptable excipient, some of the first group immunosuppressant carrier coupled operation of these synthetic nanocarriers second population with an MHC class I-restricted epitopes of one antigen-restricted epitopes coupling and / or MHC II. 在一个实施例中,该组合物处于在受试者中有效降低抗原特异性效应性T细胞的数目或活性的量。 In one embodiment, the composition is effective to reduce the amount of antigen-specific effector T cells in a subject number or activity. 优选地,当运些表位分别WMHC I类或血1C II类分子呈递时,它们是抗原特异性效应性T细胞的同源表位。 Preferably, when these epitopes are transported WMHC 1C class I or class II molecule presenting the blood, they are homologous epitope-specific effector T cells. 在一个实施例中,运些效应性T细胞的是CD4+或CD8巧细胞。 In one embodiment, the transport of these effector T cells are CD4 + or CD8 cells clever.

[0028] 在另一个方面,该组合物包含:第一群体的合成纳米载体;第二群体的合成纳米载体;W及一种可移植的移植物或治疗性蛋白;W及任选地一种药学上可接受的赋形剂,运些第一群体的合成纳米载体与免疫抑制剂偶联,运些第二群体的合成纳米载体与一种抗原的Μ肥I类限制性表位和/或MHC II类限制性表位偶联。 [0028] In another aspect, the composition comprising: a first synthetic nanocarrier groups; synthetic nanocarriers second groups; and W is a portable graft or therapeutic protein; W is and optionally one pharmaceutically acceptable excipient, some of the first population of synthetic nanocarriers immunosuppressant coupled operation of these synthetic nanocarriers second population with Μ fertilizer class I restricted epitopes of an antigen and / or MHC class II restricted epitopes conjugated. 在一个实施例中,该组合物处于在受试者中有效降低抗原特异性效应性Τ细胞的数目或活性的量。 In one embodiment, the composition is effective to reduce the amount of antigen-specific effector cells in a subject Τ number or activity. 优选地,当运些表位分别W MHC I类或MHC II类分子呈递时,它们是抗原特异性效应性Τ细胞的同源表位。 Preferably, when these epitopes are shipped W MHC class I or MHC II molecules presentation, they are homologous epitope-specific effector cells Τ. 在一个实施例中,运些效应性Τ细胞的是CD4+或CD8巧细胞。 In one embodiment, the transport of these effector cells is Τ CD4 + or CD8 cells clever.

[0029] 在另一方面,提供了一种用于治疗或预防的组合物,该组合物包含:第一群体的合成纳米载体;W及第二群体的合成纳米载体,运些第一群体的合成纳米载体与免疫抑制剂偶联,运些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联。 [0029] In another aspect, there is provided a composition for the treatment or prevention, the composition comprising: a first synthetic nanocarrier groups; and W is a second synthetic nanocarriers groups, the first group of these transport synthetic nanocarriers immunosuppressant synthesized by coupling these second population nanocarriers transported with MHC class I-restricted epitopes of an antigen and / or MHC class II restricted epitopes conjugated. 在一个实施例中,该组合物处于在受试者中有效降低抗原特异性效应性Τ细胞的数目或活性的量。 In one embodiment, the composition is effective to reduce the amount of antigen-specific effector cells in a subject Τ number or activity. 优选地,当运些表位分别WMHC I类或MHC II类分子呈递时,它们是抗原特异性效应性Τ细胞的同源表位。 Preferably, when these epitopes are transported WMHC class I or MHC II molecules presentation, they are homologous epitope-specific effector cells Τ. 在一个实施例中,运些效应性Τ细胞的是CD4+或CD8巧细胞。 In one embodiment, the transport of these effector cells is Τ CD4 + or CD8 cells clever.

[0030] 在另一方面,提供了一种组合物,该组合物包含:第一群体的合成纳米载体;W及第二群体的合成纳米载体,运些第一群体的合成纳米载体与免疫抑制剂偶联,运些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联,该组合物用于W下方法: [0030] In another aspect, there is provided a composition, the composition comprising: a first synthetic nanocarrier group; W and the second synthetic nanocarriers groups, some of the first group of transportation synthetic nanocarriers immunosuppression coupling agents, some synthetic nanocarriers shipped second population with an antigen MHC class I restricted epitope and / or class of MHC II restricted epitopes coupling, the composition is used method in W:

[0031] a.治疗,该治疗包括将所述组合物给予受试者并且在给予之前和/或之后评定受试者中的抗原特异性效应性T细胞的数目或活性的步骤; . [0031] a treatment, which treatment comprises administering to the subject the composition and prior to administration and / or after the step number or activity in a subject antigen-specific effector T cells assessed;

[0032] b.治疗,该治疗包括将所述组合物给予受试者并且在给予之前和/或之后评定受试者中的抗原特异性效应性T细胞缺失的步骤; . [0032] b treatment, which treatment comprises administering to the subject the composition of the subject and the step of antigen-specific effector T cell depletion assessed prior to administration and / or after;

[0033] C.治疗,该治疗包括根据一种治疗方案向受试者给予所述组合物的步骤,该治疗方案先前已经显示降低在一个或多个测试受试者中的抗原特异性效应性T细胞的数目或活性; [0033] C. treatment, the treatment comprises the step of administering to a subject the composition according to one treatment regimen, the treatment regimen has previously been shown to reduce antigen-specific effects in one or more subject test the number or activity of T cells;

[0034] d.使抗原特异性效应性T细胞缺失; . [0034] d of the antigen-specific effector T cell depletion;

[0035] e.如在此提供的任何运些方法中所定义的治疗或预防; . [0035] e method of any vehicle, such as those provided herein as defined in the treatment or prevention;

[0036] f.例如通过抗原特异性效应性T细胞的数目或活性的降低,对一种炎性疾病、一种自身免疫性疾病、一种过敏症、器官或组织排斥或移植物抗宿主病的治疗或预防; [0036] f., For example, by reducing the number or activity of effector antigen-specific T cells, to an inflammatory disease, an autoimmune disease, an allergic disease, organ or tissue rejection or graft versus host disease the treatment or prevention;

[0037] g.对已经经历或将要经历移植的受试者的治疗; [0037] g treatment of the subject has undergone or will undergo transplantation.;

[0038] h.对已经被给予或将要被给予一种治疗性蛋白的受试者的治疗;或 . [0038] h treatment the subject has been administered or will be administered a therapeutic protein; or

[0039] i.与一种可移植的移植物或治疗性蛋白相结合的治疗。 [0039] i. Transplantable graft with one or a combination of therapeutic protein therapy.

[0040] 在一个实施例中,该组合物处于在受试者中有效降低抗原特异性效应性T细胞的数目或活性的量。 [0040] In one embodiment, the composition is effective to reduce the amount of antigen-specific effector T cells in a subject number or activity. 优选地,当运些表位分别WMHC I类或MHC II类分子呈递时,它们是抗原特异性效应性T细胞的同源表位。 Preferably, when these epitopes are transported WMHC class I or MHC II molecules presentation, they are homologous epitope-specific effector T cells. 在一个实施例中,运些效应性T细胞是CD4+或CD8巧细胞。 In one embodiment, the transport of these effector T cells are CD4 + or CD8 cells clever.

[0041] 在另一方面,提供了一种组合物用于在在此提供的任何方法中使用的药剂的制造的用途,该组合物包含:(i)第一群体的合成纳米载体和(ii)第二群体的合成纳米载体,运些第一群体的合成纳米载体与免疫抑制剂偶联,运些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联。 [0041] In another aspect, there is provided a use for producing a composition for use in any of the methods provided herein the pharmaceutical agent, the composition comprising: Synthesis nanocarrier (i) a first population and (ii ) second population of synthetic nanocarriers, transport some synthetic nanocarriers first population immunosuppressant synthesized by coupling these second population nanocarriers transported with MHC class I-restricted epitopes of an antigen and / or MHC class II restricted epitopes conjugated. 在一个实施例中,该组合物处于在受试者中有效降低抗原特异性效应性T细胞的数目或活性的量。 In one embodiment, the composition is effective to reduce the amount of antigen-specific effector T cells in a subject number or activity. 优选地,当运些表位分别W MHC I类或MHC II类分子呈递时,它们是抗原特异性效应性T细胞的同源表位。 Preferably, when these epitopes are shipped W MHC class I or MHC II molecules presentation, they are homologous epitope-specific effector T cells. 在一个实施例中,运些效应性T细胞是CD4+或CD8巧细胞。 In one embodiment, the transport of these effector T cells are CD4 + or CD8 cells clever.

[0042] 在在此提供的任何运些组合物或用途的一个实施例中,该组合物是如所提供的任何运些方法中所定义的,和/或该组合物是供在一种治疗或预防的方法中使用,该方法包括通过静脉内、腹膜内、透粘膜、经口、皮下、肺部、鼻内、皮内或肌肉内给予来进行给予,例如, 如在所提供的任何运些方法中所定义的。 [0042] In one embodiment, any operation or use of these compositions provided herein, the composition is shipped as any of these methods are provided as defined in, and / or the composition is for use in a method of treating a method of preventing or use, the method comprises intravenous, intraperitoneal, transmucosal, oral, subcutaneous, pulmonary, intranasal, intradermal or intramuscular administration to be administered, e.g., as provided in any vehicle these methods defined.

[0043] 在另一方面,提供了一种包含在此提供的任何运些组合物的剂型。 [0043] In another aspect, there is provided a dosage form comprising any transport these compositions provided herein.

[0044] 在此提供的任何运些组合物和方法的一个实施例中,运些组合物基本上不包含B 细胞表位。 [0044] In any vehicle provided in one embodiment these compositions and methods, these compositions operation substantially does not comprise a B cell epitope.

[0045] 在此提供的任何运些组合物和方法的一个实施例中,作为包含W上提到的运些表位的蛋白质的抗原可W与运些合成纳米载体偶联。 [0045] The operation of any one embodiment herein provides compositions and methods of some embodiments, as the antigen may comprise W transported on these epitopes mentioned W and transport these proteins conjugated synthetic nanocarriers. 在另一个实施例中,包含W上提到的运些表位、但包含位于运一个或多个表位的一端或两端侧面的另外氨基酸的多肤或肤可W与运些合成纳米载体偶联。 In another embodiment, these epitopes mentioned operation comprising the W, but more or skin comprising a transport located one or both sides of the plurality of additional epitopes may be amino acid or W and skin transported some synthetic nanocarriers coupling. 在另一个实施例中,运些表位本身与运些合成纳米载体偶联。 In another embodiment, these epitopes themselves transported with the transport of these conjugated synthetic nanocarriers.

附图说明 BRIEF DESCRIPTION

[0046] 图1显示了来自Treg细胞的流式细胞检测分析的结果。 [0046] FIG. 1 shows the results from flow cytometric analysis of Treg cells.

[0047] 图2显示了使用包含免疫抑制剂(雷帕霉素或辛伐他汀)的本发明的合成纳米载体对抗原特异性的效应T细胞的数目W及FOXP3+细胞百分数的影响(在单次注射之后)。 [0047] Figure 2 shows the influence FOXP3 and synthetic nanocarriers of the present invention comprises the use of immunosuppressive agents (rapamycin or simvastatin) for antigen-specific effector T cells in the percentage of cells of W + number (in a single after injection).

[0048] 图3显示了使用包含免疫抑制剂(雷帕霉素或辛伐他汀)的本发明的合成纳米载体使順淋己结细胞的数目减少(在多次注射之后)。 [0048] Figure 3 shows synthetic nanocarriers of the present invention comprises the use of immunosuppressive agents (rapamycin or simvastatin) that the number of the cis-cyclohexyl lymph node cells decrease (after multiple injections).

[0049] 图4显示了使用包含免疫抑制剂和OVA肤的合成纳米载体的给予使CD4巧细胞和CD8巧细胞的数目减少。 [0049] FIG. 4 shows the number of synthetic nanocarrier comprising an immunosuppressant skin of OVA and administration of CD4 and CD8 cells clever clever reduction cells.

[0050] 图5显示了使用包含免疫抑制剂和OVA肤的合成纳米载体的给予使CD4巧细胞和CD8巧细胞的数目减少。 [0050] FIG. 5 shows the reduction to give the number of CD4 cells and CD8 clever clever use of synthetic nanocarriers cells comprising an immunosuppressant and a skin of OVA.

具体实施方式 Detailed ways

[0051] 在详细描述本发明之前,应当理解本发明不局限于具体举例说明的材料或工艺参数,因为运些当然可W变化。 [0051] Before describing the invention in detail, it should be understood that the present invention is not limited to the materials or process parameters specifically illustrated because these can of course be transported W changes. 还应当理解在此使用的术语只是为了描述本发明的具体实施例的目的,并不旨在限制使用可替代的术语来描述本发明。 It should also be understood that the terminology used herein is for the purpose of describing particular embodiments of the present invention, not intended to limit the use of alternative terms to describe the present invention.

[0052] 无论在前或在后,在此引用的所有公开物、专利W及专利申请,出于所有目的特此通过引用w其全文结合在此。 [0052] whether the front or rear, all publications, patents and patent applications W cited herein for all purposes in its entirety is hereby incorporated herein by reference in w.

[0053] 如在本说明书和所附权利要求中所使用,单数形式"一个(种)(a, an)"和"该(the)"包括复数指代物,除非内容另外明确表明。 [0053] As used in this specification and the appended claims, the singular forms "a (kind of) (a, an)" and "the (The)" include plural referents unless the content clearly show. 例如,提及的"一种聚合物"包括两种或更多种此类分子的混合物或单个聚合物种类的不同分子量的混合物;提及的"一种合成纳米载体"包括两种或更多种此类合成纳米载体的混合物或多个此类合成纳米载体;提及的"一种DNA分子"包括两种或更多种此类DNA分子的混合物或多个此类DNA分子;提及的"一种免疫抑制剂"包括两种或更多种此类物质的混合物或多个免疫抑制剂分子;诸如此类。 For example, "a polymer" includes mixtures of different molecular weights mentioned or mixtures of individual polymeric species of two or more such molecules; reference to "a synthetic nano-carrier" includes two or more such mixtures of synthetic nanocarriers or more such synthetic nanocarriers; "a DNA molecule" includes mixtures of two or more of such DNA molecules or a plurality of such DNA molecules mentioned; mentioned "immunosuppressant" includes mixtures of two or more such substances or more immunoreactive molecule inhibitors; like.

[0054] 如在此所使用,术语"包括(Compr i Se)"或其变体,如"包括(Compr i SeS)"或"包括(compri SingΓ意在表明包括任何列举的整体(例如,一个特点、要素、特征、特性、方法/工艺步骤或限制)或整体的组(例如,多个特点、多个要素、多个特征、多个特性、多种方法/多个工艺步骤或多个限制),但是不排除任何其他整体或整体的组。因此,如在此所使用,术语"包括"是包含性的但是不排除另外的、未列举的整体或方法/工艺步骤。 [0054] As used herein, the term "comprising (Compr i Se)" or variants thereof, such as "comprising (Compr i SeS)" or "comprise (compri SingΓ intended to indicate the entire (e.g., including any of a recited features, elements, characteristics, properties, method / process step or limitation) or group of integers (e.g., a plurality of features, a plurality of elements, a plurality of features, a plurality of characteristics, various methods / process steps or limitations plurality ), but not the exclusion of any other integer or group of integers. Thus, as used herein, the term "comprising" is inclusive of, but does not exclude additional, unrecited integers or method / process steps.

[0055] 在此处提供的任何运些组合物和方法的多个实施例中,"包括"可用"基本上由......组成(consisting essentially of)"或"由组成(consisting of)"来替代。 [0055] Examples of these transport a plurality of any of the compositions and methods of the embodiments provided herein, "comprising" Available "consisting essentially consisting ...... (consisting essentially of)" or "consisting (consisting of ) "instead. 短语"基本上由……组成"在此用来要求限定的一个或多个整体或步骤W及不实质上影响所要求的发明的特征或功能的那些。 The phrase "consisting essentially of ......" is used herein to those features or functions defined in claim one or more of W and integers or steps do not materially affect the claimed invention. 如在此所使用,术语"组成(consisting)"用来表明仅存在所列举的整体(例如,一个特点、要素、特征、特性、方法/工艺步骤或限制)或整体的组(例如,多个特点、多个要素、多个特征、多个特性、多种方法/多个工艺步骤或多个限制)。 As used herein, the term "composition (consisting)" is used to indicate the presence of only the entire recited (e.g., a feature, element, characteristic, property, method / process step or limitation) or group of integers (e.g., a plurality of characteristic, a plurality of elements, a plurality of features, a plurality of characteristics, various methods / process steps or limitations plurality).

[0化6] A.前言 [0 of 6] A. Introduction

[0057]如先前所提到的,目前常规的免疫抑制剂是广泛作用的并且通常导致免疫系统的总体系统性下调。 [0057] As previously mentioned, current conventional immunosuppressant effect is widely and generally result in an overall reduction of the systemic immune system. 在此提供的运些组合物和方法通过例如允许祀向递送至感兴趣的免疫细胞而允许更具针对性的免疫作用。 These transportation methods and compositions provided herein, for example to allow Si to be delivered to immune cells of interest to allow a more targeted by immunization. 因此,运些组合物和方法可-种更有方向性的方式实现免疫抑制。 Accordingly, these compositions and methods of operation can be - more species directional manner immunosuppression. 运可W是有帮助于减少脱祀效应和/或毒性。 Yun W may be helpful in reducing the effects de-Si and / or toxicity. 已经发现,更直接地向相关细胞(特别是APC)递送免疫抑制剂W及MHC I类限制性和/或MHC II性限制性表位能够导致效应性T细胞(例如,CD4+、CD8+T细胞)的数目和/或活性降低。 It has been found, more directly related to the cells (particularly APC) delivered immunosuppressants W and MHC class I-restricted and / or MHC II epitope can result in restriction effector T cells (e.g., CD4 +, CD8 + T cells ) number and / or activity. 效应性T细胞(即,效应T细胞)的数量和/或活性降低可W通过效应性T细胞缺失、无反应性(anergy)、效应性T细胞刺激降低或缺乏、转变成致耐受性表型等等而发生。 Number and / or activity of effector T cells (i.e., effector T cells) can be reduced by the absence of W effector T cell, anergy (anergy), effector T cell stimulating reduced or absent, into tolerogenic table type, etc. occur. 运种降低可W用若干方式评定,包括在此提供的或另外为本领域普通技术人员所知的那些。 W transported species may be reduced in several ways assessment, or those further comprising those of ordinary skill in the art provided herein. 例如,可W通过评定效应性T细胞的数目、通过评定效应性T细胞的增殖、通过评定由效应性T细胞产生的细胞因子等等来测量效应性T细胞数目或活性的降低。 For example, W can be assessed by the number of effector T cells, assessed by proliferation of effector T cells, measuring the number of effector T cells reduce the activity or the like by assessing cytokine produced by the effector T cells. 运类评定可W在体外或体内进行。 W may be transported based assessment performed in vitro or in vivo. 作为一个实例,可W对受试者给予在此提供的一种组合物并且可W在给予之后进行该测定。 As one example, W may be administered to a subject a composition provided herein and W may be determined after the administration. 运可W在从受试者获得的一个样品上完成。 W may be transported on to complete a sample obtained from the subject.

[005引如在W下实例中已经显示,本发明的组合物已经被成功地用于在体内降低效应T 细胞数目或活性。 [005 As has been shown in the cited example W, the compositions of the present invention have been successfully used in a number or activity of effector cells in vivo to reduce T. 运些实例显示通过给予包含免疫抑制剂和抗原(如OVA肤)的合成纳米载体,使CD4巧细胞和CD8巧细胞的数目W及正在分裂中的运类细胞的百分数降低。 Some examples of display operation comprising immunosuppressants and by administering an antigen (e.g., OVA skin) synthetic nanocarriers, and that the percentage of CD4 and CD8 cells clever clever class W to the number of cells dividing cells is reduced. 因此,本发明是有用的,例如,通过效应性T细胞数目和/或活性的降低来促进受试者中的致耐受性免疫应答,运些受试者例如是具有或正处于具有一种过敏症、自身免疫性疾病、一种炎性疾病、器官或组织排斥或移植物抗宿主病的风险的那些受试者。 Accordingly, the present invention is useful, e.g., to promote resistance to the immune response induced by a subject the number of effector T cells, and / or reduced activity of these transport subject having or being at, for example, to have a those subjects allergies, autoimmune disease, an inflammatory disease, organ or tissue rejection or graft-versus-host disease risk. 本发明还对促进已经经历或将要经历移植的受试者中的致耐受性免疫应答是有用的。 The present invention is also tolerant immune response induced by the subject has undergone or will undergo transplantation are useful for promotion. 本发明还有用于促进W下受试者中的致耐受性免疫应答,运些受试者已经接受、正在接受或将要接受一种治疗性蛋白,针对该治疗性蛋白,会产生或预期会产生所不希望的免疫应答。 The present invention is also used to promote tolerance induced immune responses in the subject under W, transported some subject has received, we are receiving or about to receive a therapeutic protein, the therapeutic protein against, or is expected to occur produce undesired immune response. 在一些实施例中,本发明防止或抑制了所不希望的免疫应答,运些所不希望的免疫应答可W中和某些治疗性处理的有益作用。 In some embodiments, the present invention prevents or inhibits the undesirable immune response, the transport of these unwanted immune response may be W, and the beneficial effects of certain therapeutic treatment.

[0059] 本发明的诸位发明人已经意外地并且令人惊讶地发现,通过实践在此披露的本发明,可W克服W上指出的问题和限制。 [0059] The inventors of the present invention have unexpectedly and surprisingly found that by practice of the invention disclosed herein, can be noted on the W W overcome problems and limitations. 具体地说,诸位发明人已经意外地发现,提供诱导一种致耐受性免疫应答的多种合成纳米载体组合物和有关方法是有可能的。 Specifically, the inventors have surprisingly found that, provided one kind of actuation inducing more synthetic nanocarriers compositions and related methods tolerance immune response is possible. 在多个实施例中,在此提供的方法可W用来在体外(例如,在处于培养物中的一群细胞中)或在体内(例如,在受试者中)降低效应性T细胞的数目或活性。 The number of the plurality of embodiments, the methods provided herein may be used for W in vitro (e.g., in the population of cells in culture) or in vivo (e.g., in a subject) to reduce effector T cells or activity. 虽然不希望受任何特殊理论限制,但认为在此提供的致耐受性组合物可W在体外和/或体内通过在免疫抑制剂存在下与效应性T细胞或效应性T细胞前体(例如天然T细胞)相互作用来实现效应性T细胞的数目或活性的降低。 While not wishing to be bound by any particular theory, it is believed that this provides tolerogenic compositions may be W in vitro and / or in the presence of an immunosuppressive agent by the T cell or T cell precursors effects in vivo effect (e.g. naive T cells) interacts to achieve reduced effector T cell numbers or activity.

[0060] 在此描述的运些方法包括向受试者给予一种组合物,该组合物包含:(i)第一群体的合成纳米载体和(ii)第二群体的合成纳米载体,运些第一群体的合成纳米载体与免疫抑制剂偶联,运些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联。 [0060] In operation such methods described herein include administering a composition to a subject, the composition comprising: Synthesis nanocarrier (i) a first population of the synthetic nanocarriers and (ii) a second population, some transport synthetic nanocarriers coupled with a first group of immunosuppressants, these synthetic nanocarriers shipped second population with an MHC class I-restricted epitopes of an antigen and / or MHC class II restricted epitopes conjugated. 优选地,该组合物是处于有效降低受试者中效应性T细胞数目或活性的量。 Preferably, the composition is in an amount of effector T cell numbers or activity of a subject effectively reduced. 在另一方面,提供了一种包括通过给予一种组合物而在受试者中降低效应性T细胞数目或活性的方法,该组合物包含:(i)第一群体的合成纳米载体和(ii)第二群体的合成纳米载体,运些第一群体的合成纳米载体与免疫抑制剂偶联,运些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联。 In another aspect, there is provided a method comprising a reduced number of effector T cells or activity in a subject by administering a composition of the method, the composition comprising: Synthesis nanocarrier (i) a first population and ( synthetic nanocarriers ii) a second group, the first group run some synthetic nanocarriers immunosuppressant synthesized by coupling these second population nanocarriers transported with MHC class I-restricted epitopes of an antigen and / or MHC class II restricted epitopes conjugated. 在另一方面,提供了一种包括根据一个治疗方案向受试者给予一种组合物的方法,该治疗方法先前已经显示降低在一个或多个测试受试者中的效应性T细胞数目或活性,其中该组合物包含:(i)第一群体的合成纳米载体和(ii)第二群体的合成纳米载体,运些第一群体的合成纳米载体与免疫抑制剂偶联,运些第二群体的合成纳米载体与MHC I类限制性表位和/或MHC II类限制性表位偶联。 In another aspect, there is provided a method comprising administering to the subject a composition according to one embodiment of the treatment method, the method of treatment has previously been shown to reduce the number of effector T cells in the one or more test subject or activity, wherein the composition comprises: synthesis nanocarrier (i) a first population of the synthetic nanocarriers and (ii) a second population, some synthetic nanocarriers transport coupled with a first group of immunosuppressants, these second transport synthetic nanocarriers groups with MHC class I-restricted epitopes and / or MHC class II restricted epitopes conjugated.

[0061] 还可W向在此所提供的运些受试者给予可移植的移植物、治疗性蛋白等等。 [0061] W may also be administered to the transplant operation subject these implants provided herein, the therapeutic protein and the like. 可W 在给予运些第一和第二群体的合成纳米载体之前、与此同时或在此之后向受试者给予此类组合物。 W can be transported more prior to administration of the first and second synthetic nanocarriers population, simultaneously with, or after administration of such compositions to a subject herein. 运类另外的药剂可W与或可W不与运些第一或第二群体的合成纳米载体或另一群体的合成纳米载体偶联。 Additional agents can be shipped class or W and W is not the first or second transport some synthetic nanocarriers population or another group conjugated synthetic nanocarriers. 在多个实施例中,提供的运些组合物还可W作为一个或多个维持剂量向受试者给予。 In various embodiments, there is provided a further transport these compositions as a W or more maintenance doses administered to a subject. 在此类实施例中,给予所提供的运些组合物,使得降低效应性T细胞的数目和/或活性持续一定长度的时间。 In such embodiments, the administration of these compositions transported provided, so that the time number and / or activity is reduced for a certain length of effector T cells. 在此在其他地方提供了此类长度的时间的例子。 This provides an example of such a length of time in other places.

[0062] 在又一个方面,还提供了包含运些第一群体和第二群体的合成纳米载体的运些组合物。 [0062] In a further aspect, also provides a composition comprising a transport operation some of these first groups and second population of synthetic nanocarriers. 在另一方面,提供了在此的任何运些组合物的多种剂型。 In another aspect, a variety of dosage forms in any of these compositions herein is shipped. 可W向受试者给予此类剂型,如有抗原特异性的致耐受性免疫应答(如抗原特异性的效应性T细胞数目或活性的降低)需要的受试者。 Such dosage forms can be administered to a subject W, if tolerogenic antigen-specific immune response (e.g., antigen-specific T cell number or activity of effector lowered) to a subject in need.

[0063] 在又另一个方面,提供了一种(i)产生第一群体的合成纳米载体和(ii)产生第二群体的合成纳米载体的方法,运些第一群体的合成纳米载体与免疫抑制剂偶联,运些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联。 [0063] In yet another aspect, there is provided a (i) a first population of synthetic nanocarriers and method (ii) generating a second synthetic nanocarriers generating groups, immunized with synthetic nanocarriers transport plurality of first groups inhibitor conjugate, transport of these synthetic nanocarriers second population with an MHC class I-restricted epitopes of an antigen and / or MHC class II restricted epitopes conjugated. 在一个实施例中,该方法进一步包括产生一种剂型,该剂型包含运些第一和第二群体的合成纳米载体。 In one embodiment, the method further comprises generating a dosage form comprising a first and second population of some operational synthetic nanocarriers. 在另一个实施例中,该方法进一步包括确保运些第二群体的合成纳米载体包含一种抗原的MHC I类限制性表位和/或MHC II类限制性表位。 In another embodiment, the method further comprising ensuring operation of these synthetic nanocarriers second population comprises an antigen MHC class I restricted epitope and / or MHC class II restricted epitopes. 在一个实施例中,可W通过将一种全长蛋白与运些合成纳米载体偶联来进行此种确保。 In one embodiment, one full-length proteins and transport these synthetic nanocarriers such coupling to ensure W passes. 在另一个实施例中,可W通过将包含运些表位的一种多肤或肤与运些合成纳米载体偶联来进行此种确保。 In another embodiment, W may be transported by these epitope comprising a multi-op skin or skin with some conjugated to synthetic nanocarriers ensure such. 在此其他地方提供了多种用于测试合成纳米载体W便确定可W产生的免疫应答的方法。 Elsewhere herein provides a method for testing a variety of immune synthetic nanocarriers W W will determine response generated. 优选地,运些组合物有效于降低效应性T细胞的数目或活性。 Preferably, these compositions effectively transported to reduce the number of effector T cells or activity. 此类方法连同本领域普通技术人员已知的其他方法可W用来确保运些合成纳米载体与所希望的表位偶联。 Such methods together with other methods known to those of ordinary skill in the art may be used to ensure W transported with some synthetic nanocarriers desired epitope conjugates.

[0064] 在再另一个实施例中,该方法进一步包括对使用一种组合物或剂型的效应性T细胞数目和/或活性的降低进行评定,该组合物或剂型包含运些第一群体和第二群体的合成纳米载体。 [0064] In yet another embodiment, the method further comprises the use of a composition or dosage form, or the number of effector T cells decreased and / or activity to be assessed, the composition or dosage form comprises a plurality of first groups and transport synthetic nanocarriers second population. 在又另一个实施例中,该方法进一步包括制造一种包含运些第一群体和第二群体的合成纳米载体的组合物、或可供受试者给予的剂型。 In yet another embodiment, the method further comprises producing a composition comprising a plurality of first groups and the second operation group of synthetic nanocarriers, subject, or a dosage form for administration.

[0065] 在另一方面,还提供了通过在此提供的任何方法产生的运些组合物或剂型。 [0065] In another aspect, also provides some transport composition or dosage form produced by any of the methods provided herein.

[0066] 现在,在W下将更详细地描述本发明。 The present invention [0066] Now, described in more detail in W.

[0067] B.定义 [0067] B. defined

[006引"给予(Administering)"或"给予(administration)"意指W药理学上有用的方式向受试者提供一种物质。 [006 cited "administration (Administering)" or "administration (Administration)" means the W pharmacologically useful way to provide a substance to the subject.

[0069] "过敏原"是在受试者中可W引起一种所不希望的(例如,1型超敏性)免疫应答(即,过敏性应答或反应)的任何物质。 [0069] "allergen" is caused in a subject that may be one kind of W is not desirable (e.g., type 1 hypersensitivity) immune response (i.e., response or allergic reaction) any substance. 过敏原包括但不限于:植物过敏原(例如,花粉、豚草过敏原)、昆虫过敏原、昆虫叮咬过敏原(例如,蜜蜂叮咬过敏原)、动物过敏原(例如,宠物过敏原,如动物皮屑或猫Fei dl抗原)、乳胶过敏原、霉菌过敏原、真菌过敏原、化妆品过敏原、 药物过敏原、食物过敏原、灰尘、昆虫毒液、病毒、细菌,等等。 Allergens include but are not limited to: plant allergens (e.g., pollens, ragweed allergens), insect allergens, insect bites allergens (e.g., bee stings allergens), animal allergens (e.g., pet allergens, such as animal Fei dl or cat dander antigen), latex allergens, mold allergens, fungal allergens, cosmetic allergens, drug allergens, food allergens, dust, insect venom, viruses, bacteria, and so on. 食物过敏原包括,但不限于:牛奶过敏原、鸡蛋过敏原、坚果过敏原(例如,花生或树坚果过敏原,等等(例如,胡桃、腰果,等等))、鱼过敏原、贝类过敏原、大豆过敏原、豆类过敏原、种子过敏原、W及小麦过敏原。 Food allergens include, but are not limited to: milk allergen, egg allergen, nut allergens (eg, peanuts or tree nut allergens, etc. (eg, walnut, cashew, etc.)), allergens fish, shellfish allergens, soy allergens, bean allergens, seed allergens, W and wheat allergens. 昆虫叮咬过敏原包括作为或与蜜蜂叮咬、胡蜂叮咬、马蜂叮咬、黄蜂叮咬等等相关联的过敏原。 Insect sting or as allergens include bee sting, bites wasps, wasp stings, bites wasp allergens like associated. 昆虫过敏原还包括屋尘蛾过敏原(例如,Der P1过敏原及蜂卿过敏原。药物过敏原包括作为或与抗生素、NSAID、麻醉剂等等相关联的抗原。花粉过敏原包括草过敏原、树过敏原、 杂草过敏原、花过敏原,等等。可W用在此提供的任何运些组合物和方法对发展或正处于发展针对在此提供的任何运些过敏原的一种所不希望的免疫应答的风险的受试者进行治疗。 可W用提供的任何运些组合物和方法治疗的受试者还包括已经对或正处于对提供的任何运些过敏原过敏的风险的那些受试者。 Insect allergens further comprising a house dust moth allergen (e.g., Der P1 allergens and bee Qing allergens. Drug allergens include associated as or antibiotics, NSAID is, anesthetics and other antigens. Pollen allergens include grass allergen, the one kind of tree allergens, weed allergens, flowers allergens, and so on. W can be transported with any of these compositions and methods are provided herein for development or are in development for any transportation provided herein some allergens the risk of undesirable immune response of the subject treated. these transport any subject composition and method can be provided with a W treatment already comprises or further allergens are at risk for any of these transport offers those subjects.

[0070] 在此又称为"过敏性病症"的"过敏"是其中对一种物质存在所不希望的(例如,1型超敏性)免疫应答(即,过敏性应答或反应)的任何病症。 [0070] Here, also known as "allergic disorder" and "allergy" in which the presence of undesirable substance (e.g., type 1 hypersensitivity) immune response (i.e., response or allergic reaction) any disease. 此类物质在此被称为过敏原。 Such substances are referred to herein allergens. 过敏或过敏性病症包括但不限于:过敏性哮喘、花粉症、等麻疹、湿疹、植物过敏、蜜蜂叮咬过敏、 宠物过敏、乳胶过敏、霉菌过敏、化妆品过敏、食物过敏、过敏性鼻炎或鼻伤风、局部过敏性反应、过敏症、特应性皮炎、超敏反应、W及其他过敏性病症。 Allergy or allergic disorders include, but are not limited to: allergic asthma, hay fever, such as measles, eczema, plant allergies, bee sting allergies, pet allergies, latex allergies, mold allergies, cosmetics allergy, food allergy, allergic rhinitis or nasal cold , local allergic reactions, anaphylaxis, atopic dermatitis, hypersensitivity, W, and other allergic disorders. 该过敏性反应可W是针对任何过敏原的一种免疫反应的结果。 The allergic reaction may be the result of any W against allergens an immune response. 在一些实施例中,该过敏症是一种食物过敏。 In some embodiments, the allergy is a food allergy. 食物过敏包括但不限于:牛奶过敏、鸡蛋过敏、坚果过敏、鱼过敏、贝类过敏、大豆过敏或小麦过敏。 Including but not limited to food allergies: milk allergy, egg allergy, nut allergy, allergic to fish, shellfish allergy, soy allergy or wheat allergy.

[0071] 在用于向受试者给予的一种组合物或剂型的背景下的"有效的量"是指在该受试者中产生一个或多个所希望的免疫应答(例如,产生一种致耐受性免疫应答,例如CD8巧细胞的增殖、活化、诱导、募集的降低)的该组合物或剂型的量。 [0071] In the context of a composition or dosage form for administration to a subject of an "effective amount" refers to an immune response to one or more of the desired subject (e.g., to produce a the amount of the composition or dosage form seed tolerogenic immune responses, such as proliferation of CD8 cells Coincidentally, activation induced, decreased recruitment) of. 因此,在一些实施例中,有效的量是产生一种或多种运些所希望的免疫应答的在此提供的一种组合物的任何量。 Thus, in some embodiments, an effective amount to produce an amount of one or more of any of these operation desired a composition provided herein immune response. 运个量可W出于体外或体内目的。 A transport amount W for purposes of in vivo or in vitro. 对于体内目的而言,该量可W是临床医师将认为对有抗原特异性耐受需要的受试者可W具有临床益处的量。 For in vivo purposes, the amount of W is clinician that the subject antigen-specific tolerance required amount of W may have clinical benefit.

[0072] 有效的量可W仅设及降低所不希望的免疫应答的水平,虽然在一些实施例中,它设及完全防止所不希望的免疫应答。 [0072] The effective amount may be provided only W and the reduced level of undesirable immune response, although in some embodiments, and it is disposed completely prevent undesired immune responses. 有效的量还可W设及延迟所不希望的免疫应答的发生。 Occurrence of an immune response an effective amount of delay and provided W may not be desirable. 有效的量还可W是产生所希望的治疗终点或所希望的治疗结果的在此提供的一种组合物的量。 W is also an amount effective to produce the desired therapeutic result amount of a composition provided herein or therapeutic endpoint desired. 有效的量优选在受试者中导致针对抗原的致耐受性免疫应答。 Effective amount preferably results in an immune response against induced tolerance to an antigen in a subject. 可W通过常规方法来监测任何上述免疫应答的实现。 W may be used to monitor any immune response to achieve the above by a conventional method. 优选地,在此有效的量是例如通过效应性T细胞缺失而导致效应性T细胞数目和/或活性降低的那些量。 Preferably, in this example, an effective amount of effector T cells by deletion resulting in the number of effector T cells, and / or reduced activity of those quantities.

[0073] 在提供的任何运些组合物和方法的一些实施例中,该有效的量是其中使所希望的免疫应答在该受试者中持续至少1周、至少2周、至少1个月、至少2个月、至少3个月、至少4个月、至少5个月、至少6个月、至少9个月、至少1年、至少2年、至少5年、或更长时间的量。 [0073] In some embodiments of any transport these compositions and methods provided, wherein the effective amount is that the desired immune response in the subject for at least 1 week, at least 2 weeks, at least 1 month , at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 9 months, at least 1 year, at least 2 years, at least 5 years, or longer amount. 在提供的任何运些组合物和方法的其他实施例中,该有效的量是产生一种可测量的所希望的免疫应答(例如,可测量的免疫应答(例如,针对一种特异性抗原)降低)持续至少1周、至少2 周、至少1个月、至少2个月、至少3个月、至少4个月、至少5个月、至少6个月、至少9个月、至少1年、至少2年、至少5年、或更长时间的量。 In other embodiments of any transport these compositions and methods provided herein, the effective amount is desired to produce a measurable immune response (e.g., a measurable immune response (e.g., directed to a specific antigen) reduction) for at least one week, at least 2 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 9 months, at least 1 year, at least 2 years, at least five years, or longer amount.

[0074] 当然,有效的量将取决于正被治疗的具体受试者;病症、疾病或素乱的严重性;个体患者的参数,包括年龄、身体状况、体型W及体重;治疗持续时间;同步疗法(如果有)的性质;特定的给药途径、W及健康执业医师的知识和专业技能内的类似因素。 [0074] Of course, the effective amount will depend on the particular subject being treated; condition, disease or disorder severity prime; individual patient parameters including age, physical condition, size and weight W; duration of the treatment; nature synchronization therapy (if any); the particular route of administration, W and similar factors within the health practitioner knowledge and expertise. 运些因素为本领域普通技术人员所熟知,并且可W仅用常规实验方法解决。 These factors known to those of ordinary transport skill in the art, and W can be resolved with only routine experimentation. 通常优选的是使用最大剂量,即根据正确医学判断的最高安全剂量。 It is generally preferred to use the maximum dose, the highest safe dose according to sound medical judgment. 然而,本领域普通技术人员应当理解,出于医学原因、 屯、理原因或几乎任何其他原因,患者可W坚持较低剂量或耐受剂量。 However, those of ordinary skill in the art will appreciate that, for medical reasons, Tuen, almost physical reasons or for any other reason, the patient may insist W lower dose or tolerable dose.

[0075] 通常,在本发明组合物中的免疫抑制剂和/或抗原的剂量可W在从约10yg/kg至约100,000yg/kg的范围内。 [0075] Generally, an immunosuppressive agent in the compositions of the present invention and / or W in a dose of antigen can range from about 100,000yg 10yg / kg to about / kg of. 在一些实施例中,运些剂量可W在从约0.1 mg/kg至约lOOmg/kg的范围内。 In some embodiments, W can be transported more doses in the range from about 0.1 mg / kg to a range of about lOOmg / kg of. 在再其他的实施例中,运些剂量可W在从约0.1 mg/kg至约25mg/kg、约25mg/kg至约50mg/kg、约50mg/kg至约7 5mg/kg、或约7 5mg/kg至约1 OOmg/kg的范围内。 In still other embodiments, transport of these doses may be W is from about 0.1 mg / kg to about 25mg / kg, to about 25mg / kg to about 50mg / kg, to about 50mg / kg to about 7 5mg / kg, or from about 7 5mg / kg in the range of about 1 OOmg / kg of. 可替代地,可W基于提供希望的量的免疫抑制剂和/或抗原的合成纳米载体的数目给予该剂量。 Alternatively, W may be provided based on the number desired amount of immunosuppressant and / or synthetic nanocarriers the dose administered antigen. 例如,有用的剂量包括每剂量大于1〇6个、1〇7个、1〇8个、1〇9个或10"个合成纳米载体。有用的剂量的其他例子包括每剂量从约1 X 1〇6至约1 X l〇w个、约1 X 107至约1 X 109个、或约1 X 108至约1 X 1〇9 个合成纳米载体。 For example, useful doses include doses greater than 1〇6 each one, a 1〇7, 1〇8 months, or 10 1〇9 "synthetic nanocarriers. Other examples of useful doses per dose comprises from about 1 X 1 to about 1 X l〇w 〇6, about 1 X 107 to about 1 X 109, or about 1 X 108 to about 1 X 1〇9 synthetic nanocarrier.

[0076] "抗原"意指B细胞抗原或T细胞抗原。 [0076] An "antigen" is meant a B cell antigen or T cell antigen. "抗原的一种或多种类型"意指共享相同的、 或基本上相同的抗原特性的分子。 "One or more types of antigens" means share the same, or a molecule substantially identical antigenic properties. 在一些实施例中,抗原可W是蛋白质类、多肤类、肤类、月旨蛋白类、糖脂类、多核巧酸类、多糖类,或被包含或表达在细胞中。 In some embodiments, the antigen can be proteins W, multi skin type, skin type, purpose month proteins, glycolipids, Qiao polynuclear acids, polysaccharides, or contained or expressed in a cell. 在一些实施例中,例如当没有很好定义或表征运些抗原时,运些抗原可W被包含在细胞或组织制品、细胞碎片、细胞外来体、条件培养基等等之内。 In some embodiments, for example when there is no well defined or characterized transported more antigens, these antigens may transport W is contained within a cell or tissue preparation, cell debris, cell exosomes, conditioned medium and so forth. 可W与受试者暴露于引起所不希望的免疫应答者相同的形式将一种抗原与运些合成纳米载体组合,但该抗原还可W是其片段或衍生物。 And W may be exposed to the same subject in the form of an immune response by causing undesirable to transport one antigen with some synthetic nanocarriers compositions, but also W is the antigen fragment or derivative thereof. 然而,当为一个片段或衍生物时,针对由运样的受试者遇到的形式的所希望的免疫应答是使用所提供的运些组合物和方法的优选结果。 However, when a fragment or derivative thereof, for form encountered by the sample transport subject an immune response is desired, a preferable result the compositions and methods of use provided by some operation.

[0077] "抗原特异性"是指由该抗原或其部分的存在引起的、或产生特异性地识别或结合该抗原的多种分子的任何免疫应答。 [0077] "antigen-specific" means that caused by the presence of the antigen or a portion thereof, or any binding immunospecifically recognizes or more molecules of the antigen response. 例如,在该免疫应答是抗原特异性抗体产生的情况下, 产生了特异性地结合该抗原的多种抗体。 For example, in the case where the immune response is an antigen-specific antibody production, produce a variety of antibody specifically binds the antigen. 作为另一个实例,在该免疫应答是抗原特异性的CD8+T细胞或CD4+T细胞增殖和/或活性的情况下,该增殖和/或活性可W由单独地或与MHC 分子复合的该抗原或其部分的识别引起。 As another example, the immune response is a CD8 + CD4 + T cell proliferation at antigen-specific T cells or and / or activity of the situation, the proliferation and / or activity may be alone or W is complexed with the MHC molecule antigen recognition or a portion thereof caused.

[0078] 与在此提供的一种疾病、素乱或病症"相关的抗原"是可W针对该疾病、素乱或病症、作为其结果、或与其结合而产生所不希望的免疫应答的抗原;可W产生该疾病、素乱或病症(或其症状或影响)的病因的抗原;和/或可W产生作为该疾病、素乱或病症的症状、结果或影响的所不希望的免疫应答的抗原。 [0078] and provided herein a disease, disorder or condition prime "associated antigen" is W against the disease, disorder or condition element, as a result of, or in connection with an antigen to produce an undesired immune response ; W can produce the disease, disorder or condition prime cause (or a symptom or effect) an antigen; and / or W as an immune response to the disease, disorder or condition symptoms hormone, result or effect undesirable antigen. 优选地,在一些实施例中,于在此提供的运些组合物和方法中使用一种与疾病、素乱或病症等相关的抗原将会导致针对该抗原和/或细胞(该抗原由运些细胞表达、在运些细胞之上或之中表达)的致耐受性免疫应答。 Preferably, in some embodiments, use of a disease, disorder, or other disorder associated prime antigen to transport these compositions and methods provided herein will result against the antigen and / or cells (the antigen by operational these cells express, on the expression of these cells or in operation) tolerogenic immune response. 运些抗原可W处于与具有该疾病、素乱或病症的受试者中所表达的相同形式,但还可W是其片段或衍生物。 These antigens may be transported in the same form as W having the disease, disorder or condition of the subject Su as expressed, but also is W fragment or derivative thereof. 然而,当为一个片段或衍生物时,针对运一受试者所遭遇的运种形式的一种希望的免疫应答是使用所提供的运些组合物和方法的优选结果。 However, when a fragment or derivative thereof, a hope for a transport operation forms the subject an immune response encountered is preferred that the composition and method of operation using these offer. 在一些实施例中,与疾病、素乱或病症相关的运些抗原包含MHC I类限制性表位和/或MHC II类限制性表位。 In some embodiments, associated with a disease, disorder or condition of the prime operation comprising these MHC class I restricted antigen epitopes and / or MHC II-restricted epitopes. 在一些实施例中,运些抗原基本上不包含B细胞表位,如当该疾病、素乱或病症是自身免疫性疾病或过敏并且包括该B细胞表位将使所不希望的免疫应答加剧时。 In some embodiments, these antigens do not substantially contain transported B cell epitope, such as when the disease, disorder or condition is hormone autoimmune disease or allergies comprising an epitope and the B cell immune response would intensify undesirable Time. 在其他实施例中,运些抗原不包含B细胞表位。 In other embodiments, the antigen does not contain more transport B-cell epitopes.

[0079] 在一个实施例中,该抗原是与一种炎性疾病、自身免疫性疾病、器官或组织排斥、 或移植物抗宿主病相关联的抗原。 [0079] In one embodiment, the antigen is an inflammatory disease, an autoimmune disease, organ or tissue rejection, or graft versus host disease associated antigen. 此类抗原包括:自身抗原,如髓憐脂碱性蛋白、胶原(例如,11型胶原)、人软骨甜39、嗜铭粒蛋白A、甜130-RAPS、蛋白脂质蛋白、纤维蛋白、核蛋白、 核仁蛋白(例如,小核仁蛋白)、甲状腺刺激因子受体、组蛋白、糖蛋白gp 70、核糖体蛋白、丙酬酸脱氨酶、脱氨硫辛酷胺乙酷转移酶、毛囊抗原、人原肌球蛋白同工型5、线粒体蛋白、膜腺β-细胞蛋白、髓銷少突胶质细胞糖蛋白、膜岛素、谷氨酸脱簇酶(GAD)、谷蛋白、W及其片段或衍生物。 Such antigens include: self-antigens, such as aliphatic marrow pity basic protein, collagen (e.g., collagen type 11), human cartilage sweet 39, Ming addicted grain protein A, sweet 130-RAPS, proteolipid protein, fibrin, nuclear proteins, nucleolar proteins (e.g., small nucleolar protein), thyroid stimulating factor receptor, histone, glycoprotein gp 70, ribosomal protein, propionic acid deaminase pay deamination lipoic cool cool transferases amine acetate, follicle antigen, human tropomyosin isoform 5, mitochondrial proteins, cell membrane proteins β- gland, medullary pin oligodendrocyte glycoprotein, membrane Insulin, cluster enzyme glutamate release (of GAD), gluten, W and fragments or derivatives thereof. W下在表1中提供了其他的自身抗原。 W provides the other autoantigens in Table 1.

[0080] 抗原还可W包括与器官或组织排斥相关联的那些抗原。 [0080] W antigens also include those antigens associated with organ or tissue rejection. 此类抗原的实例包括,但不限于:来自同种异体细胞的抗原,例如来自一种同种异体细胞提取物的抗原;和来自其他细胞的抗原,如内皮细胞抗原。 Examples of such antigens include, but are not limited to: antigens from allogeneic cells, for example, an antigen from allogeneic cell extract; and antigens from other cells, such as endothelial cell antigen.

[0081] 抗原还包括与一种过敏相关联的那些抗原。 [0081] The antigen-associated antigens also include those with one allergy. 此类抗原包括在此其他地方描述的运些过敏原。 Such antigens include transport some allergens described elsewhere herein.

[0082] 抗原还包括与一种可移植的移植物相关联的那些抗原。 [0082] antigens also include those graft antigens associated with a portable. 此类抗原与一种可移植的移植物、或具有一种可移植的移植物的接受者中的一种所不希望的免疫应答相关联,该所不希望的免疫应答由于将该可移植的移植物引入到该接受者中而产生,该可移植的移植物可W被呈递用于由免疫系统的细胞识别并且该可移植的移植物可W产生一种所不希望的免疫应答。 Such antigens with a portable graft, or a portable receiver having one graft in an undesirable immune response associated with a undesirable immune response because the portable introduced into the graft recipient is generated, the portable W graft may be presented to the immune system by the cell identification and the portable W graft can produce one kind of immune response is undesirable. 移植抗原包括与器官或组织排斥或移植物抗宿主病相关联的那些抗原。 Transplantation antigens including organ or tissue rejection or graft versus host disease that is associated antigens. 可W从一种生物材料的细胞中或从与一种可移植的移植物相关的信息中获得或衍生出移植抗原。 Or W may be obtained or derived from transplantation antigens information associated with a portable cellular grafts from a biological material. 移植抗原通常包括蛋白质类、多肤类、肤类、脂蛋白类、糖脂类、多核巧酸类、多糖类,或被包含或表达在细胞中。 Transplantation antigens typically comprises proteins, multi-skin type, skin type, lipoproteins, glycolipids, Qiao polynuclear acids, polysaccharides, or contained or expressed in a cell. 与一种可移植的移植物相关的信息是关于可W用来获得或衍生出移植抗原的一种可移植的移植物的任何信息。 Associated with a portable information about the graft W may be used to obtain any information derived from one kind or transplantation antigens may be transplanted graft. 此类信息包括关于将要预期会存在于一种可移植的移植物的细胞之中或之上的抗原的信息,例如像序列信息;抗原和/或它们的MHC I、MHC II或B细胞呈递限制的类型或种类。 Such information includes information regarding expected to be present in the cell in a portable or on graft antigen, such as sequence information; antigen and / or their MHC I, MHC II restricted presentation or B cell the type or kind. 此类信息还可W包括关于W下各项的信息:可移植的移植物的类型(例如,自身移植物、同种异体移植物、异种移植物)、该移植物的分子组成和细胞组成、该移植物源自其中或该移植物有待被移植至其上的身体位置(例如,全部或部分的器官、皮肤、骨骼、神经、腫、神经元、血管、脂肪、角膜等等)。 Such information may also include information regarding the W at W: Types of graft transplants (e.g., autograft, allograft, xenograft), graft composition of the molecules and cells, or wherein the graft from the graft is to be transplanted to a body location (e.g., all or part of an organ, skin, bone, nerve, swelling, neurons, blood vessels, fat, cornea, etc.) on its.

[0083] 抗原还包括与一种治疗性蛋白相关联的抗原,运些抗原可W被呈递用于由免疫系统的细胞识别并且运些抗原可W产生针对该治疗性蛋白的一种所不希望的免疫应答。 [0083] The further antigen comprises a therapeutic protein associated antigen, W can be transported more antigen-presenting cells for recognition by the immune system and these antigens may be transported W to produce a therapeutic protein for which the undesired immune response. 治疗性蛋白抗原通常包括蛋白质类、多肤类、肤类、脂蛋白类,或被包含在或表达在细胞之中、之上或由细胞表达。 Therapeutic protein antigens typically comprises proteins, multi-skin type, skin type, lipoproteins, or contained within the cell or expressed in, on or expressed by a cell.

[0084] 抗原可W是完全定义或表征的抗原。 [0084] W is the antigen may be fully characterized or defined antigen. 然而,在一些实施例中,抗原不是完全定义或表征的。 However, in some embodiments, the antigen is not fully defined or characterized. 因此,抗原还包括被包含在细胞或组织制品、细胞碎片、细胞外来体或条件培养基内的那些抗原,并且在一些实施例中可W处于此种形式递送。 Thus, antigens include those antigens are further contained within a cell or tissue preparation, cell debris, or foreign cell conditioned medium, and in some embodiments, W can be delivered in this form.

[0085] "评定一种免疫应答"是指对体外或体内免疫应答的水平、存在或不存在、降低、增加等等进行的任何测量或测定。 [0085] "Evaluation of an immune response" refers to the level of immune response in vitro or in vivo, the presence or absence, decrease, increase in any measured or determined for the like. 可W在从受试者中获得的一个或多个样品上进行此类测量或测定。 W can be measured or determined in such one or more sample obtained from the subject. 可W用在此提供的任何方法或本领域中已知的其他方法来进行此类评定。 W may be used any of the methods provided herein or other methods known in the art to assess such.

[0086] "处于风险中"的受试者是健康执业医师认为其具有患上在此所提供的疾病、素乱或病症的机会的受试者、或健康执业医师认为其具有经历在此所提供的所不希望的免疫应答的机会的受试者。 [0086] "at risk" of a subject is health practitioners believe it has suffered in the opportunity offered by this disease, disorder or condition of the prime subjects, or health practitioner believes it has the experience in this subjects the chance of immune response provided by undesirable.

[0087] "自身免疫性疾病"是其中免疫系统针对自身(例如,一个或多个自身抗原)激发一种所不希望的免疫应答的任何疾病。 [0087] "Autoimmune disease" against which the immune system itself (e.g., one or more self antigens) stimulate one kind of any disease undesired immune response. 在一些实施例中,自身免疫性疾病包含作为自身祀向免疫应答的一部分的对身体细胞的异常破坏。 In some embodiments, the autoimmune disease comprises the destruction of abnormal cells as part of the body's own immune response to worship. 在一些实施例中,自身破坏表现为一个器官(例如,结肠或膜腺)的功能失常。 In some embodiments, the performance of self-destruction of an organ (e.g., colon or membrane gland) dysfunctional. 在此其他地方描述了自身免疫性疾病的实例。 Here is described elsewhere examples of autoimmune diseases. 本领域普通技术人员已知另外的自身免疫性疾病,并且本发明在此方面不受限制。 It is known to those of ordinary skill in another autoimmune disease, and the present invention is not limited in this respect.

[0088] 如在此所使用的"平均值"是指算术平均值,除非另外指出。 [0088] As the "average value" as used herein refers to the arithmetic mean, unless otherwise indicated.

[0089] "B细胞抗原"意指触发B细胞中的免疫应答的任何抗原(例如,由B细胞或其上的受体特异性地识别的抗原)。 [0089] "B cell antigen" is meant any antigen B cells triggers an immune response (e.g., B cell receptor, or by a specifically recognizes the antigen). 在一些实施例中,作为T细胞抗原的抗原也是一种B细胞抗原。 In some embodiments, the T cell antigen as an antigen but also a B cell antigen. 在其他实施例中,该T细胞抗原并不也是一种B细胞抗原。 In other embodiments, the T cell antigen is not also a B cell antigen. B细胞抗原包括但不限于蛋白质类、 肤类、小分子、W及碳水化合物。 B cell antigens include, but are not limited to proteins, skin type, small molecules, W, and carbohydrates. 在一些实施例中,B细胞抗原包括非蛋白抗原(即,不是一种蛋白质或肤抗原)。 In some embodiments, B cell antigen comprises a non-protein antigen (i.e., not a protein antigen, or skin). 在一些实施例中,该B细胞抗原包含一种自身抗原。 In some embodiments, the B cell antigen comprises one autoantigen. 在其他实施例中,该B 细胞抗原从一种过敏原、自身抗原、治疗性蛋白、或可移植的移植物中获得或衍生出。 In other embodiments, the B cell antigen from an allergen, an autoantigen, therapeutic protein, or transplant grafts obtained or derived.

[0090] "同时"意指W时间相关的、优选地时间充分相关的方式向受试者给予两种或更多种物质,W便提供对免疫应答的调节。 [0090] "simultaneously" means that the time-dependent W, sufficient time is preferably related embodiment the administration of two or more substances to a subject, will provide W regulation of the immune response. 在多个实施例中,可W通过W相同剂型给予两种或更多种物质而发生同时给予。 In various embodiments, administration may occur simultaneously W administration of two or more substances in the same dosage form W. 在其他实施例中,同时给予可W涵盖W不同剂型、但在指定时间段内、优选在1个月内、更优选在1周内、再更优选在1天内并且甚至更优选在1小时内给予两种或更多种物质。 In other embodiments, W encompasses W may be administered simultaneously in different dosage forms, but in one hour within the specified period, preferably within one month, more preferably within 1 week, still more preferably within one day, and even more preferably administration of two or more substances.

[0091] "偶联(Couple)"或"偶联(Coupled)"或"偶联(Couples)"(诸如此类)意指使一个实体(例如一个部分)与另一个实体在化学上结合。 [0091] "coupling (Couple)" or "coupling (Coupled)" or "coupling (Couples)" (like) means that one entity (e.g., a portion) binds chemically with another entity. 在一些实施例中,偶联是共价的,运意味着该偶联是在两个实体之间在存在共价键的背景下发生的。 In some embodiments, the coupling is covalent, meaning that the coupling operation takes place between the two entities in the presence of a background of a covalent bond. 在非共价的实施例中,通过非共价相互作用介导非共价偶联,运些非共价相互作用包括但不限于:电荷相互作用、亲和相互作用、金属配位、物理吸附、主-客体相互作用、疏水相互作用、ττ堆积相互作用、氨键相互作用、范德华相互作用、磁相互作用、静电相互作用、偶极-偶极相互作用、和/或其组合。 In an embodiment of the non-covalent, by non-covalent interactions mediated by non-covalent coupling, non-covalent interactions of these operation include, but are not limited to: charge interactions, affinity interactions, metal coordination, physical adsorption , host - guest interactions, hydrophobic interactions, ττ stacking interactions, ammonia bonding interactions, van der Waals interactions, magnetic interactions, electrostatic interactions, dipole - dipole interactions, and / or combinations thereof. 在多个实施例中,封装是偶联的一种形式。 In various embodiments, the package is one form of coupling.

[0092] "衍生的"意指由一种材料或与一种材料相关的信息制备的,而不是从该材料"获得的"。 [0092] "derived from" means made from one material or information associated with one material, instead of the material from "acquired." 此类材料可W是直接取自一种生物材料的材料的实质性地改变或加工的形式。 Such materials may be in the form of substantially W is changed or processed directly from a biological material material. 此类材料还包括从与一种生物材料相关的信息所产生的材料。 Such materials also include materials related to the information generated from a biological material.

[0093] "剂型"意指在一种适用于向受试者给予的介质、载体、媒介物或装置中的药理学和/或免疫学活性材料。 Medium, carrier, vehicle or means [0093] "dosage form" means suitable for administration to a subject in a pharmacologically and / or immunologically active materials.

[0094] "封装"意指将物质的至少一个部分封闭在合成纳米载体内。 [0094] "Package" means at least a portion of the material is enclosed within the synthetic nanocarrier. 在一些实施例中,一种物质被完全封闭在合成纳米载体内。 In some embodiments, a substance is completely enclosed within the synthetic nanocarrier. 在其他实施例中,被封装的物质的大部分或全部不暴露于该合成纳米载体外的局部环境。 In other embodiments, most or all of the synthesis is not exposed to the outside of the local environment nanocarrier encapsulated substance. 在其他实施例中,不超过50%、40%、30%、20%、 10%或5% (重量/重量)暴露于局部环境。 In other embodiments, no more than 50%, 40%, 30%, 20%, 10% or 5% (wt / wt) are exposed to the local environment. 封装不同于吸附,吸附是将物质的大部分或全部置于合成纳米载体的表面上,并且使得该物质暴露于该合成纳米载体外的局部环境。 Unlike the package, and the adsorption is synthesized on the surface of the nanocarrier most or all substance is placed, and so that the substance is exposed to the outside of the synthetic nanocarrier local environment.

[00Μ]"表位",又称为抗原决定簇,是由免疫系统,确切地由例如抗体、Β细胞、或Τ细胞识别的抗原部分。 [00Μ] "epitope", also known as antigenic determinants by the immune system, for example, an antibody specifically, Beta cells, or cells recognize antigens Τ portion. 如在此所使用,"MHC I类限制性表位"是通过在有核细胞上发现的MHC I类分子呈递给免疫细胞的表位。 As used herein, "class I MHC restricted epitope" by MHC class I molecules on nucleated cells found in an epitope is presented to immune cells. II类限制性的表位"是通过在抗原呈递细胞(APC)上(例如,在专口的抗原呈递免疫细胞上,如在巨隧细胞、B细胞W及树突状细胞上,或在非造血细胞如肝细胞上)发现的MHC II类分子呈递给免疫细胞的表位/'B细胞表位"是由抗体或B细胞识别的分子结构。 Class II restricted epitope "by antigen presenting on (the APC) (e.g., the E I antigen-presenting cells on an immune cell, such as a tunnel on the giant cells, B cells and dendritic cells W, or non- hematopoietic cells such as liver cells), MHC II molecules found in the form of epitope presented to the immune cells / 'B cell epitope "is a molecular structure of an antibody or a B-cell recognition. 在一些实施例中,该表位本身是一种抗原。 In some embodiments, the epitope is an antigen itself. 优选地,为了产生在此提供的所希望的致耐受性免疫应答,MHC I类限制性表位和/或MHC II类限制性表位是当WMHC I类或血1C II类分子呈递时结合细胞上的受体并且导致效应性T细胞的缺失、活性抑制等等的那些表位。 Preferably, in order to produce the desired herein provided tolerogenic immune response, of MHC class I restricted epitope and / or MHC II-restricted epitope is a binding when WMHC class I or class II molecule presenting the blood 1C receptors on cells and leads to absence of effector T cells, inhibiting activity like those epitopes. 由于效应性T细胞分泌细胞因子并且会导致所不希望的免疫应答的其他分子, 那么运些细胞的减少可W下调其他所不希望的免疫应答。 Since effector T cells secrete cytokines and other molecules can lead to undesired immune response, then the operation of these cells can be reduced down W other undesirable immune response.

[0096]许多表位是本领域普通技术人员已知的,并且根据本发明的一些方面适合的示例性表位包括但不限于列在免疫表位数据库(WWW, immuneepitope.org,维塔· R(Vita R)、札瑞布斯基· L(Zarebski L)、格林鲍姆· JA(Greenbaum JA)、埃马米· Η巧mami H)、霍夫· I 化oof I)、萨利姆.N(Salimi N)、达美乐.R(Damle R)、赛特*A(Sette A)、彼得斯·Β (Peters Β),免疫表位数据库2.0(The immune epitope database 2.0).核酸研究(Nucleic Acids Res) ,2010年1月;38(数据库发行号):D854-62;该免疫表位数据库的全部内容W及2011年8月的IEDB 2.4版本的所有数据库条目,并且具体地说在其中披露的所有表位通过引用结合在此)中的那些。 [0096] Many epitopes are known to those of ordinary skill in the art, and include, but are not limited to a column in the Immune Epitope database (WWW, immuneepitope.org, Vita · R in accordance with some aspects of the invention suitable exemplary epitope (Vita R), 札瑞布斯基 · L (Zarebski L), Greenbaum · JA (Greenbaum JA), Ai Mami · Η Qiao mami H), Hough · I of oof I), Salem. N (Salimi N), Domino .R (Damle R), Dorset * A (Sette A), Peters · Β (Peters Β), immune epitope database 2.0 (The immune epitope database 2.0). nucleic Acids Research (nucleic Acids Res), 2010 Jan; 38 (database issue number): D854-62; the entire contents of all database entries in the immune epitope database and August 2011 W of IEDB 2.4 version, and in particular where disclosure All epitopes incorporated herein by reference) those. 还可W使用公开可获得的算法来鉴定表位,运些算法例如在W下文献中描述的算法:王平(Wang P)、西德尼· J(Sidney J)、金姆· Y化im Y)、赛特-A(SetteA)、伦德.(KLund 0)、尼耳森·Μ(Μθΐ3Θη M)、彼得斯.B^eters B),2010, 用于HLA DR、DPW及DQ分子的肤结合预测(peptide binding predictions for HLA DR,DP and DQ mole州les),BMC生物信息学(BMC Bioinformatics)2010,ll:568;王平(Wang P)、 西德尼· J(Sidney J)、道· C(Dow C)、莫斯· B(Mothe B)、赛特· A(Sette A)、彼得斯· B (Peters B),2008,MHC II类肤结合预测的系统性评定和一致方法的评估(A systematic assessment of MHC class II peptide binding predictions and evaluation of a consensus approach),化oS计算生物学(PLoS Comput Biol.)4(4):e100 0048;尼耳森·Μ (Nielsen Μ)、伦德-0(Lund 0),2009,顺-比对,用于MHC II类版结合预测的基于人工神经网络白勺比对算、法(NN-al ign .An artificial neural network-based W may also be used publicly available algorithms to identify epitopes, for example, described in the operation of these algorithms W algorithm in the literature: Ping (Wang P), Sidney · J (Sidney J), Kim · Y im of Y), Dorset -A (SetteA), Lund. (KLund 0), Nigeria ear Mori · Μ (Μθΐ3Θη M), Peters .B ^ eters B), 2010, for HLA DR, DPW and DQ molecules skin binding prediction (peptide binding predictions for HLA DR, DP and DQ mole states les), BMC Bioinformatics (BMC Bioinformatics) 2010, ll: 568; Ping (Wang P), Sidney · J (Sidney J), Road · C (Dow C), Moss · B (Mothe B), Dorset · a (Sette a), Peters · B (Peters B), 2008, MHC class II binding skin systematic assessment and prediction methods consistent evaluation (a systematic assessment of MHC class II peptide binding predictions and evaluation of a consensus approach), computational biology of oS (PLoS Comput Biol.) 4 (4): e100 0048; Mori Nigeria ear · Μ (Nielsen Μ), Lund -0 (Lund 0), 2009, cis - comparing, for MHC class II binding prediction based version of artificial neural network operator than white spoon, France (NN-al ign .An artificial neural network-based alignment algorithm for MHC class II peptide binding prediction),BMC生物信息学(BMC Bioinformatics),10: 296;尼耳森· M(Nielsen M)、伦德高· CXLundegaard C)、伦德· 0 化und 0) ,2007,使用新颖的稳定化矩阵比对方法SMM-比对预测MHC II类结合亲和性(Prediction of MHC class II binding affinity using SMM-align,a novel stabilization matrix alignment method),BMC生物信息学(BMC Bioinformatics),8: 238;裴·ΗΗ(Βιιί HH)、西德尼.KSidney J)、彼得斯·Β(ΡθΐθΓ8 B)、萨塞默塞·Μ (Sathiamui'thy Μ)、真一· A(Sinichi A)、潘顿· KA(Purton ΚΑ)、莫斯· BR(Mo化e BR)、奇萨利FV)、沃特金斯-OUWatkins DI)、赛特.A(Sette A),2005,免疫遗传学(Immunogene tics),57:304-314;斯图尔尼奥洛· T(Sturniolo Τ)、博诺· E(Bono E)、 下· J(Ding J)、控德里查尼· L(Raddrizzani L)、图雷西· 0(Tuereci 0)、萨因· U(Sahin U)、布雷桑勒《KBraxenthaler Μ)、加利亚西·Ρ(63ΐΐ3ζζΐ F)、普罗狄 alignment algorithm for MHC class II peptide binding prediction), BMC Bioinformatics (BMC Bioinformatics), 10: 296; 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• Larson of arsen Qiao Qiao), Lund (KLund 0), Nepal ear Sen .KNielsen M), 2006, an improved method (improved method for predicting linear B-cell epitopes) linear epitope prediction for B cells, immunity (Immunome Res) 2: 2; Ponomarenko · JV (Ponomarenko JV), Byrne · PE (Bourne PE), 2007, antibodies - protein interaction: a reference data set and the predictive evaluation tools (Antibody- protein inter said ctions: benchmark datasets and prediction tools ev lu said said tion), BMC structural Biology (BMC Struct Biol ) seven sixty-four; * [rho] Anderson haast of aste Andersen P), Nigeria ear Mori · Μ (Nielsen Μ), of Lund · 0 und 0), 2006, using a protein 3D structure prediction discontinuous B cell epitopes the residue (Prediction of residues in discontinuous B-cell epitopes using protein 3D structures), protein Science (protein Sci) 15: 2558-2567; Ponomarenko · JV (Ponomarenko JV), PEI · H (Bui H), Li · W (Li W), Fu Side · N (Fusseder N)、伯恩•阳(Bourne PE)、赛特Sette A、彼得斯· B^eters B),2008,ElliP;ro:用于预测抗体表位的新型基于结构的工具(ElliPro:曰new structure-based tool for the prediction of 曰ntibody epitopes), BMC 生物信息学(BMC Bioinformatics)9:514;尼耳森.M(Nielsen M)、伦德高.C (Lundegaard C)、布里加·Τ(Β1;ΐχ1ΐθ;Γ T)、彼得斯《B^eters B)、赛特》A(Sette A)、贾斯特逊《SGustesen S)、布斯·8(Βιιιΐ8 S)W及伦德《(KLund 0),2008,化〇5计算生物学(PLoS Comput Biol.)4(7)e100 0107,肤结合具有已知序列的任何HLA-DR分子的定量预测: NetMHCIIpan(Quantitative predictions of peptide binding to any HLA-DR mole州le of known seq uence:NetMHCIIpan);运些文献各自的全部内容通过引用结合在此W用于披露用于鉴定表位的方法和算法。 (Fusseder N), Bern • male (Bourne PE), Dorset Sette A, Peters · B ^ eters B), 2008, ElliP; ro: a novel epitope prediction-based tool structure (ElliPro: said new structure-based tool for the prediction of said ntibody epitopes), BMC Bioinformatics (BMC Bioinformatics) 9: 514; Sen Nepal ear .M (Nielsen M), Sor high .C (Lundegaard C), Brigadeiro · Τ (Β1; ΐχ1ΐθ; Γ T), Peters "B ^ eters B), Dorset" A (Sette A), Chester Johnson "SGustesen S), Booth · 8 (Βιιιΐ8 S) W and Lund" ( KLund 0), 2008, Computational Biology of 〇5 (PLoS Comput Biol) 4 (7) e100 0107, any skin quantitative binding of HLA-DR molecules having a known sequence prediction:. NetMHCIIpan (quantitative predictions of peptide binding to any HLA-DR mole states le of known seq uence: NetMHCIIpan); Yun entire contents of each of these documents is incorporated herein by reference, discloses a method and W algorithm for epitope identification.

[0097] 在此提供的表位的其他实例包括如SEQ ID NOs :1-943所提供的MHC I类限制性表位、MHC II类限制性表位W及B细胞表位中的任何。 [0097] Other examples of epitope as provided herein include SEQ ID NOs: any MHC Class I-restricted epitope provided 1-943, MHC II-restricted epitopes W and B cell epitopes. 不希望受任何具体理论束缚,MHC I类限制性表位包括在SEQ ID NO: 1-186中列出的那些,MHC II类限制性表位包括在SEQ ID NO: 187-537中列出的那些,并且B细胞表位包括在SEQ ID NO:538-943中列出的那些。 Without wishing to be bound by any particular theory, MHC I-restricted epitope comprises SEQ ID NO: those, MHC II-restricted epitope comprising 1-186 listed in SEQ ID NO: 187-537 are listed in the those, and B cell epitope comprises SEQ ID NO: 538-943 as those listed. 运些表位包括MHC I类限制性自身抗原、过敏原的MHC II类限制性表位、W及自身抗原和过敏原的B 细胞表位。 These epitope includes transport MHC class I-restricted self-antigens, allergens MHC class II restricted epitopes, W, and B-cell epitopes of autoantigens and allergens.

[0098] "产生"意指本身直接地或间接地(如但不限于,通过依赖某人的语言或行为采取行动的不相关的第Ξ方)引起一种行为(如一种免疫应答(例如,致耐受性免疫应答))发生。 [0098] "generate" means itself directly or indirectly (such as, but not limited, by the action of a person dependent language or behavior unrelated second party Ξ) causes an action (e.g., an immune response (e.g., tolerogenic immune response)) occurs.

[0099] "鉴定"是允许临床医师将受试者识别为可能从在此提供的运些方法和组合物中获益者的任何行为或一系列行为。 [0099] "Identification" is the subject to allow the clinician to identify any possible behavior beneficiaries from some transport methods and compositions provided herein, or a series of acts. 优选地,鉴定的受试者是对如在此所提供的致耐受性免疫应答有需要者。 Preferably, the subject is identified immune tolerance induced as provided herein answer those in need. 该行为或一系列行为可W是本身直接地或间接地,如但不限于,通过依赖某人的语言或行为采取行动的不相关的第Ξ方。 The act or series of acts may be W is itself directly or indirectly, such as, but not limited to, the action by relying on one's language or behavior of the Ξ party irrelevant.

[0100] "免疫抑制剂"意指引起APC具有免疫抑制性(例如,致耐受性作用)的一种化合物。 [0100] "immunosuppressive agent" is meant having caused APC (e.g., a tolerogenic role) of a compound of immune suppression. 免疫抑制作用通常是指由APC产生或表达细胞因子或其他因子,该产生或表达降低、抑制或防止所不希望的免疫应答、或促进所希望的免疫应答。 Immunosuppression generally refers to the production or expression of cytokines or other factors by the APC, the production or expression reduce, inhibit or prevent undesired immune responses, or to promote the desired immune response. 当该APC对识别由该APC呈递的抗原的免疫细胞产生一种免疫抑制作用时,该免疫抑制作用被称为是对呈递的抗原特异的。 When the APC of the immune cells to recognize antigen presented by the APC produce an immunosuppressive effect, which is known to be immunosuppressive effect of the presentation of antigen specific. 此种作用在此还称为致耐受性作用。 Such action also referred to herein tolerogenic effect. 不受任何具体理论束缚,认为该免疫抑制或致耐受作用是该免疫抑制剂被递送至该APC上的结果,优选在一种抗原(例如,一种给予的抗原或已经存在于体内的抗原)存在的情况下。 Not to be bound by any particular theory, that the effect of immunosuppressive or tolerogenic immune inhibitor is delivered to the results on the APC, preferably in an antigen (e.g. a given antigen or antigens present in the body have been ) in the presence of. 因此,该免疫抑制剂包括提供针对一种抗原的致耐受性免疫应答的化合物,运些化合物可W被或可W不被提供在相同的组合物或不同的组合物中。 Thus, the immunosuppressive agent comprises a compound providing tolerogenic immune response against one antigen, W may be transported or these compounds may be W is not provided in the same composition or different compositions. 在一个实施例中,该免疫抑制剂是引起APC促进一个或多个免疫效应细胞中的调节表型的免疫抑制剂。 In one embodiment, the immunosuppressive agent is one or more promoting APC induced immune effector cells immunosuppressant regulating phenotype. 例如,可W通过抑制抗原特异性CD4巧细胞或CD8巧细胞的产生、诱导、刺激或募集;Treg细胞(例如,CD4+CD25hi曲化XP3+化eg细胞)的产生、诱导、刺激或募集等等,对该调节性表型进行表征。 For example, W can be produced by antigen-specific CD4 or CD8 cells coincidence coincidence cell inhibition, induction, stimulation or recruitment; generating Treg cells (e.g., CD4 + CD25hi eg of curvature of XP3 + cells), inducing, stimulating, or the like to raise , the regulatory phenotype characterization. 运可W是CD4巧细胞或CD8巧细胞或B细胞转变为调节性表型的结果。 W can be transported clever CD4 or CD8 cells or B-cell coincidence results into regulatory phenotype. 运还可W是在其他免疫细胞(如CD8巧细胞、巨隧细胞W及iNKT细胞)中诱导化xP3的结果。 W may also be transported in other immune cells (e.g., CD8 cells Qiao, W and giant cells tunnel iNKT cells) results in the induction of xP3. 在一个实施例中,该免疫抑制剂是在APC加工抗原之后影响该APC的应答的免疫抑制剂。 In one embodiment, the immunosuppressive agent is an immunosuppressive influence the response of APC APC after antigen processing. 在另一个实施例中,该免疫抑制剂不是干扰抗原加工的免疫抑制剂。 In another embodiment, the immunosuppressive agent is not interfere with antigen processing immunosuppressant. 在一个另外的实施例中,该免疫抑制剂不是一种细胞调亡信号分子。 In a further embodiment, the immunosuppressive agent is not an apoptosis signaling molecule. 在另一个实施例中,该免疫抑制剂不是憐脂。 In another embodiment, the immunosuppressive agent is not pity fat.

[0101] 免疫抑制剂包括但不限于:抑制素;mTOR抑制剂,如雷帕霉素或一种雷帕霉素类似物;TGF-β信号剂;TGF-β受体激动剂;组蛋白去乙酷化酶抑制剂,如曲古抑菌素A (Trichostatin A);皮质类固醇类;线粒体功能抑制剂,如鱼藤酬(rotenone) ;P38抑制剂; NF-κβ抑制剂,如她io、地塞米松(Dexamethasone)、TCPA-1、IKK VII;腺巧受体激动剂;前列腺素E2激动剂(PGE2),如米索前列醇(Misoprostol);憐酸二醋酶抑制剂,如憐酸二醋酶4抑制剂(PDE4),如咯利普兰(Rolipram);蛋白酶体抑制剂;激酶抑制剂;G-蛋白偶联受体激动剂;G-蛋白偶联受体括抗剂;糖皮质激素类;类视黄醇类;细胞因子抑制剂;细胞因子受体抑制剂;细胞因子受体活化剂;过氧化物酶体增殖物激活受体括抗剂;过氧化物酶体增殖物激活受体激动剂;组蛋白去乙酷化酶抑制剂;巧调憐酸酶(calcineurin)抑制剂;憐酸酶抑制剂;PI3邸抑制剂,如TGX-221; [0101] immunosuppressive agents include but are not limited to: inhibin; of mTOR inhibitors such as rapamycin or one rapamycin analogues; TGF-β signal reagent; TGF-β receptor agonists; histone cool b inhibitors, such as trichostatin A (trichostatin A); corticosteroids; mitochondrial function inhibitors such as rotenone pay (rotenone); P38 inhibitors; NF-κβ inhibitors, as she IO, dexamethasone (dexamethasone), TCPA-1, IKK VII; Qiao adenovirus receptor agonist; prostaglandin E2 agonist (of PGE2), such as misoprostol (misoprostol); pity acid esterase inhibitors, such as acid pity 4 diacetate enzyme inhibitors (PDE4), such as rolipram (of rolipram); proteasome inhibitors; kinase inhibitors; G-protein coupled receptor agonists; G-protein coupled receptor antagonist comprising; glucocorticoids hormones; retinoids; cytokine inhibitors; inhibitors of cytokine receptors; cytokine receptor activator; peroxisome proliferator-activated receptor antagonist comprising; peroxisome proliferator-activated receptor agonist; histone acetyl cool inhibitors; clever modulation pity glucuronidase (calcineurin) inhibitor; pity acid inhibitor; Di a PI3 inhibitors, such as TGX-221; 自隧(autophagy)抑制剂,如3-甲基腺嚷岭;芳香控受体抑制剂;蛋白酶体抑制剂I(PSI); W及氧化的ATP,如P2X受体阻断剂。 Since tunneling (autophagy in) inhibitors, such as 3-methyl gland cried ridge; aromatase inhibitor control receptor; proteasome inhibitor I (PSI); W and oxidized ATP, such as P2X receptor blockers. 免疫抑制剂还包括ID0、维生素D3、环抱霉素类(如环抱霉素A)、芳香控受体抑制剂、白襲芦醇、硫挫嚷岭(Aza)、6-琉基嚷岭(6-MP)、6-硫鸟嚷岭(6-TG)、FK506、萨菲菌素A(sanglifehrin A)、沙美特罗(salmeterol)、吗替麦考酪醋(mycophenolate mofetil) (MMF)、阿司匹林W及其他C0X抑制剂、尼氣酸、雌Ξ醇W及雷公藤甲素(triptolide)。 Further comprising the ID0 immunosuppressant, vitamin D3, cyclosporine (such as cyclosporine A), an aromatic control receptor inhibitors, white passage resveratrol, sulfur frustrated cried ridge (Aza), 6- thiol cried ridge (6 -MP), 6- sulfur birds cried ridge (6-TG), FK506, sanglifehrin A (sanglifehrin A), salmeterol (salmeterol), mycophenolate casein vinegar (mycophenolate mofetil) (MMF), aspirin W C0X and other inhibitors, Nigeria acid gas, and W female Ξ alcohol triptolide (triptolide). 在多个实施例中,免疫抑制剂可W包含在此提供的任何药剂(agent)。 In various embodiments, the immunosuppressive agent may comprise any agent W (Agent) provided herein.

[0102] 免疫抑制剂可W是对APC直接提供免疫抑制(例如,致耐受)作用的一种化合物,或该免疫抑制剂可W是间接(即,在给予之后W某种方式被加工之后)提供免疫抑制(例如,致耐受)作用的一种化合物。 After [0102] W is an immunosuppressive agent can be directly APC immunosuppression (e.g., tolerogenic) acting compound, or immunosuppressant which W may be indirect (i.e., after administration of the workpiece W in some way ) providing immunosuppressive compound (e.g., tolerogenic) action. 因此,免疫抑制剂包括在此提供的任何化合物的前药形式。 Accordingly, prodrug forms of immunosuppressive agents comprising any of the compounds provided herein.

[0103] 免疫抑制剂还可W包括编码在此提供的产生免疫抑制(例如,致耐受)免疫应答的肤、多肤或蛋白质的核酸。 [0103] W may further comprise an immunosuppressant immune suppression (e.g., tolerogenic) provided herein skin encoding an immune response, multiple skin protein or nucleic acid. 因此,在多个实施例中,免疫抑制剂是编码一种产生免疫抑制(例如,致耐受)免疫应答的肤、多肤或蛋白质的核酸,并且该免疫抑制剂是与合成纳米载体偶联的核酸。 Thus, in various embodiments, for generating coded immunosuppressant immunosuppressive (e.g., tolerogenic) immune response of skin, skin plurality protein or nucleic acid, and the immunosuppressive agent is conjugated with a synthetic nanocarriers nucleic acid.

[0104] 该核酸可W是DNA或RNA,如mRNA。 [0104] W is the nucleic acid can be DNA or RNA, such as mRNA. 在多个实施例中,运些发明的组合物包含一种互补物,如一种全长互补物,或在此提供的任何运些核酸的一种简并物(由于遗传密码的简并性)。 In various embodiments, these compositions transported to the invention comprises one complement, such as one full-length complement thereof, or any of these transport provided herein and was a degenerate nucleic acid (due to the degeneracy of the genetic code) . 在多个实施例中,核酸是一种表达载体,当转染至细胞系中时,该表达载体可W被转录。 In various embodiments, the nucleic acid is an expression vector, when transfected into cell lines, the expression vector can be transcribed W. 在多个实施例中,该表达载体可W包含除其他之外的质粒、逆转录病毒、或腺病毒。 In various embodiments, the adenoviral vector expressing W may contain in addition other plasmid, retrovirus, or. 可W 使用标准的分子生物学方法来分离或合成核酸,例如通过使用聚合酶链反应来产生核酸片段,然后对该核酸片段进行纯化并且克隆至表达载体中。 W may be using standard molecular biology methods for nucleic acid isolated or synthesized, for example, nucleic acid fragments is generated by using the polymerase chain reaction, nucleic acid fragments and then purified and cloned into the expression vector. 对本发明的实践有用的另外技术可W在2007年约翰威立国际出版公司(John Wil巧and Sons,Inc.)的现代分子生物学实验方案(Current Protocols in Molecular Biology);分子克隆:实验室手册(第Ξ版)萨姆布鲁克(Molecular Cloning : A Laboratory Manual (Third Edition) Joseph Sambrook),彼得麦卡勒姆癌症研究所(Peter MacCallum Cancer Institute),墨尔本(Me化ourne),澳大利亚(Aus化alia);大卫罗素(David Russell),德克萨斯大学西南医学中屯、(University of Texas Southwestern Medical Center),达拉斯(Dallas),冷泉港(Cold Spring 化rbor)中找到。 Useful for the practice of the invention may additionally technology in 2007, John Wiley & W International Publishing Company (John Wil clever and Sons, Inc.) Of modern molecular biology protocols (Current Protocols in Molecular Biology); Molecular Cloning: A Laboratory Manual (Ξ first edition), Sambrook (Molecular Cloning: A Laboratory Manual (Third Edition) Joseph Sambrook), Peter MacCallum cancer Institute (Peter MacCallum cancer Institute), Melbourne (Me of ourne), Australia (Aus of alia ); David Russell (David Russell), University of Texas southwestern Medical Tuen, (University of Texas southwestern Medical Center), Dallas (Dallas), Cold Spring Harbor (Cold Spring of rbor) found.

[0105] 在多个实施例中,在此提供的运些免疫抑制剂与合成纳米载体偶联。 [0105] In various embodiments, the operation of these immunosuppressants synthetic nanocarriers conjugate provided herein. 在多个优选的实施例中,免疫抑制剂是除了构成该合成纳米载体的结构的材料之外的一种成分。 In preferred embodiments, the immunosuppressive agent is an ingredient in addition to the structure of the material constituting the synthetic nanocarrier. 例如, 在一个实施例中,在合成纳米载体由一种或多种聚合物构成的情况下,该免疫抑制剂是除了运一种或多种聚合物之外的并且与其偶联的一种化合物。 For example, in one embodiment, in the case of synthetic nanocarriers consisting of one or more polymers, which is an immunosuppressant compound in addition to the operation of one or more polymers and coupled thereto . 作为另一个实例,在一个实施例中,在该合成纳米载体由一种或多种脂质构成的情况下,该免疫抑制剂又是除了运一种或多种脂质之外的并且与其偶联的。 As another example, in one embodiment, in a case where the synthetic nanocarriers made of one or more lipids, the immunosuppressant is other than addition of one or more lipid transport and coupling thereto linked. 在多个实施例中,例如在该合成纳米载体的材料也产生一种免疫抑制(例如,致耐受)作用的情况下,该免疫抑制剂是产生一种免疫抑制(例如, 致耐受)作用的、除了该合成纳米载体材料之外而存在的一种成分。 In various embodiments, for example, to produce a material of the immunosuppression synthetic nanocarriers (e.g., tolerogenic) action case, the immunosuppressive agent is an immunosuppressive generated (e.g., tolerogenic) effect, one component in addition to the support material of the synthetic nano-exist.

[0106] 其他示例性的免疫抑制剂包括但不限于:小分子药物、天然产物、抗体(例如,对抗CD20、CD3、CD4的抗体)、基于生物制剂的药物、基于碳水化合物的药物、纳米颗粒、脂质体、 RMi、反义核酸、适体、氨甲蝶岭、NSAID;芬戈莫德(fingolimod);那他珠单抗(natalizumab);阿仑单抗(alemtuzumab);抗-CD3;他克莫司(tac;rolimus)(FK506)等等。 [0106] Other exemplary inhibitors include, but are not limited to immunization: small molecule drugs, natural products, antibodies (e.g., against CD20, CD3, antibodies to CD4), based on bio-pharmaceutical agents, carbohydrate-based drugs, nanoparticles , liposomes, RMi, antisense, aptamer, methotrexate Ridge, NSAID; fingolimod (fingolimod); natalizumab (natalizumab); alemtuzumab (alemtuzumab); anti--CD3; tacrolimus (tac; rolimus) (FK506), and so on. 另外的免疫抑制剂对于本领域的技术人员是已知的,并且本发明在此方面不受限制。 Additional immunosuppressive agent for those skilled in the art are known, and the present invention is not limited in this respect.

[0107] "炎性疾病"意指其中发生所不希望的炎症的任何疾病、素乱或病症。 [0107] "Inflammatory disease" means any disease where inflammation occurs undesirably, hormone disorder or condition.

[0108] 免疫抑制剂或抗原的"负载"是基于整个合成纳米载体中的材料的总重量,与合成纳米载体偶联的该免疫抑制剂或抗原的量(重量/重量)。 [0108] immunosuppressants or an antigen "load" is a material based on total weight of the entire synthetic nanocarriers, the amount of the immunosuppressive agent or antigen conjugated with synthetic nanocarriers (wt / wt). 通常,该负载被计算为在一个群体的合成纳米载体上的平均值。 Typically, the load is calculated as an average over a group of synthetic nanocarriers of. 在一个实施例中,平均在运些第一群体的合成纳米载体上的免疫抑制剂的负载是在0.0001%与50%之间。 In one embodiment, the average load on the synthetic immunosuppressant nanocarrier operation of some of the first group is between 0.0001% and 50%. 在另一个实施例中,平均在该第一和/或第二群体的合成纳米载体上的该抗原的负载是在0.0001 %与50%之间。 In another embodiment, the average load on the antigen synthetic nanocarriers of the first and / or second population is between 0.0001% and 50%. 在又一个实施例中,该免疫抑制剂和/或抗原的负载是在0.01%与20%之间。 In yet another embodiment, the immunosuppressive agent and the load / or antigen is between 0.01 and 20%. 在一个另外的实施例中,该免疫抑制剂和/或抗原的负载是在0.1%与10%之间。 In a further embodiment, the immunosuppressive agent and the load / or antigen is between 0.1% and 10%. 在再一个另外的实施例中,该免疫抑制剂和/或抗原的负载是在1 %与10%之间。 In yet a further embodiment, the immunosuppressive agent and the load / or antigen is between 1% and 10%. 在又另一个实施例中,平均在一个群体的合成纳米载体上的该免疫抑制剂和/或该抗原的负载是至少0.1 %、至少0.2%、至少0.3%、至少0.4%、至少0.5%、至少0.6%、至少0.7%、至少0.8%、至少0.9%、至少1 %、至少2%、至少3%、至少4%、至少5%、至少6%、至少7%、至少8%、至少9%、至少10%、至少11 %、至少12%、至少13%、至少14%、至少15%、至少16%、至少17 %、至少18%、至少19%、或至少20%。 In yet another embodiment, the average in the population of a synthetic nanocarriers the immunosuppressive and / or load the antigen is at least 0.1%, at least 0.2%, at least 0.3%, at least 0.4%, at least 0.5%, at least 0.6%, at least 0.7%, at least 0.8%, at least 0.9%, at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9 %, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, or at least 20%. 在又一个另外的实施例中,平均在一个群体的合成纳米载体上的该免疫抑制剂和/或该抗原的负载是0.1%、0.2%、0.3%、0.4%、0.5%、0.6%、0.7%、0.8%、0.9%、1%、2%、3%、4%、 5%、6%、7%、8%、9%、10%、11%、12%、13%、14%、15%、16%、17%、18%、19%、或20%。 In yet a further embodiment, the immunosuppressive agent and / or loading of the antigen on average a synthetic nanocarriers group was 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7 %, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20%. 在W上运些实施例中的一些实施例中,平均在一个群体的合成纳米载体上的该免疫抑制剂和/或该抗原的负载不大于25%。 W transported in some embodiments some embodiments, the immunosuppressive agent and / or the load on the synthetic antigen average nanocarrier a group of not more than 25%. 在多个实施例中,如实例中所描述对该负载进行计算。 In multiple embodiments, as described in the examples of the calculation load.

[0109] 在提供的任何运些组合物和方法的多个实施例中,可W如下计算该负载:收集近似3mg的合成纳米载体,并且离屯、W使上清液与合成纳米载体沉淀物分离。 Collected approximately 3mg synthetic nanocarriers, and from the village, W nanocarriers the supernatant and the precipitate was synthesized: [0109] embodiment, W may be calculated as follows any of the plurality of load transport these compositions and methods provided in the embodiments separation. 向该沉淀物添加乙腊,并且对样品进行超声处理并离屯、W去除任何不溶的材料。 To the precipitate was added acetic wax, and the samples were sonicated and centrifuged Tun, W remove any insoluble material. 在RP-HPLC上注射该上清液和沉淀物,并且在278nm处读取吸光度。 The supernatant and pellet injection on RP-HPLC, and the absorbance was read at 278nm. 该沉淀物中的yg用来计算%包载(负载),在上清液和沉淀物中的yg用来计算回收的总yg。 The precipitate is used to calculate the% entrapped yg (load), yg supernatant and the precipitate is collected to calculate the total yg.

[0110] "维持剂量"是指在初始剂量已经在受试者中产生免疫抑制(例如,致耐受)应答之后,为了维持所希望的免疫抑制(例如,致耐受)应答而向受试者给予的剂量。 After [0110] "maintenance dose" is one which has been produced immunosuppression (e.g., tolerogenic) response in a subject at an initial dose, immune suppression in order to maintain a desired (e.g., tolerogenic) in reply to the test They were given doses. 维持剂量例如可W是维持在该初始剂量之后所实现的致耐受作用、防止该受试者中的所不希望的免疫应答、或防止该受试者变为处于经历所不希望的免疫应答(包括所不希望的水平的免疫应答) 风险的受试者的剂量。 Maintenance dose is maintained for example W tolerisation After this initial dose is achieved, the subject to prevent undesirable immune responses, or prevent an immune response in the subject becomes undesirably subjected (not including the desired level of immune response) dose subjects at risk. 在一些实施例中,该维持剂量是足W维持适当水平的所希望的免疫应答的剂量。 In some embodiments, the maintenance dose is sufficient to maintain an appropriate level W of the desired dose of an immune response. 在一些实施例中,该维持剂量是足W维持适当水平的CD4巧细胞或CD8巧细胞数目或活性、或抵抗用导致所不希望的免疫应答的一种物质(agent)进行的激发的剂量。 In some embodiments, the maintenance dose is sufficient to maintain an appropriate level W of CD4 or CD8 cells Qiao Qiao cell numbers or activity, or resistance resulted in a dose excited with a substance (Agent) undesired immune response performed.

[0111] "合成纳米载体的最大尺寸"意指沿着该合成纳米载体的任何轴线测量的纳米载体的最大尺寸。 [0111] The maximum size of the nanocarrier means measured along any axis of the synthetic nanocarriers "maximum dimension of a synthetic nanocarrier." "合成纳米载体的最小尺寸"意指沿着该合成纳米载体的任何轴线测量的合成纳米载体的最小尺寸。 "Synthesis nanocarrier minimum size" means the minimum dimension along the axis of a synthetic nanocarrier any measurement of the synthetic nanocarrier. 例如,对于球形合成纳米载体而言,合成纳米载体的最大和最小尺寸将是基本上相同的,并且将是该合成纳米载体的直径的大小。 For example, for spherical synthetic nanocarriers, the maximum and minimum dimensions of a synthetic nanocarrier will be substantially the same, and the size of the diameter of the synthetic nanocarriers. 类似地,针对立方形合成纳米载体,合成纳米载体的最小尺寸将是它的高度、宽度或长度中最小的,而合成纳米载体的最大尺寸将是它的高度、宽度或长度中最大的。 Similarly, for the synthesis of cubic nanocarriers, synthetic nanocarrier minimum dimension will be its height, width or length of the smallest, and the maximum size of the synthetic nanocarriers will be its height, width or length of the largest. 在一个实施例中,基于样品中的合成纳米载体的总数,样品中至少75 %、优选地至少80 %、更优选地至少90 %的运些合成纳米载体的最小尺寸等于或大于lOOnm。 In one embodiment, the total number of synthetic nanocarriers in the sample, the sample based on at least 75%, preferably at least 80%, more preferably at least 90% of the minimum dimension of these op synthetic nanocarriers equal or greater than lOOnm. 在一个实施例中,基于样品中的合成纳米载体的总数,样品中至少75%、优选地至少80%、更优选地至少90%的运些合成纳米载体的最大尺寸等于或小于5 μπι。 In one embodiment, the total number of synthetic nanocarriers in the sample, the sample based on at least 75%, preferably at least 80%, more preferably at least 90% of the largest dimension of the operation of these synthetic nanocarriers less than or equal to 5 μπι. 优选地,基于样品中的合成纳米载体的总数,样品中至少75%、优选地至少80%、更优选地至少90%的运些合成纳米载体的最小尺寸大于110皿、更优选地大于120皿、更优选地大于130加1、并且还更优选地大于150加1。 Preferably, the total number of synthetic nanocarriers in the sample, the sample based on at least 75%, preferably at least 80%, the minimum size of at least 90% of the transported synthetic nanocarriers more preferably more than 110 dishes, more preferably greater than 120 dish , more preferably greater than 130 plus 1, and further more preferably greater than 150 plus 1. 发明的合成纳米载体的最大尺寸与最小尺寸的长宽比可W取决于实施例而变化。 Synthetic nanocarriers largest dimension to the smallest dimension of the invention, the aspect ratio W may vary depending on the embodiment. 例如,运些合成纳米载体的最大尺寸与最小尺寸的长宽比可^从1:1至1,〇〇〇,〇〇〇:1、优选地从1:1至1〇〇,〇〇〇:1、更优选地从1:1至1〇,〇〇〇:1、更优选地从1:1至1000:1、再更优选地从1:1至100:1、并且又更优选地从1:1至10:1变化。 For example, some synthetic nanocarriers transport the largest dimension to the smallest dimension of the aspect ratio can be from 1 ^: 1-1, 〇〇〇, 〇〇〇: 1, preferably from 1: 1 to 1〇〇, 〇〇〇 : 1, more preferably from 1: 1 to 1〇, 〇〇〇: 1, more preferably from 1: 1 to 1000: 1, still more preferably from 1: 1 to 100: 1, and still more preferably from 1: 1 to 10: 1 variation. 优选地,基于样品中的合成纳米载体的总数,样品中至少75%、优选地至少80%、更优选地至少90%的运些合成纳米载体的最大尺寸等于或小于3皿、更优选地等于或小于2μπι、更优选地等于或小于Ιμπι、更优选地等于或小于80化m、更优选地等于或小于600nm、并且还更优选地等于或小于500nm。 Preferably, the total number of synthetic nanocarriers in the sample, the sample based on at least 75%, preferably at least 80%, more preferably at least 90% of the operational maximum size of these synthetic nanocarriers 3 or less dish, more preferably equal to or less 2μπι, more preferably equal to or less than Ιμπι, more preferably of equal to or less than 80 m, more preferably equal to or less than 600nm, and still more preferably equal to or less than 500nm. 在多个优选的实施例中,基于样品中的合成纳米载体的总数,样品中的至少75%、优选地至少80%、更优选地至少90%的运些合成纳米载体的最小尺寸等于或大于lOOnm、更优选地等于或大于120nm、更优选地等于或大于130nm、更优选地等于或大于14化m、并且还更优选地等于或大于150nm。 In preferred embodiments, the total number based on the minimum size of the synthetic nanocarriers in the sample, the sample is at least 75%, preferably at least 80%, more preferably at least 90% of these synthetic nanocarriers transport or greater lOOnm, more preferably equal to or greater than 120nm, more preferably equal to or greater than 130nm, more preferably equal to or greater than of 14 m, and still more preferably equal to or greater than 150nm. 通过将运些合成纳米载体悬浮在一种液体(通常是水性)介质中,并且使用动态光散射(DLS)(例如使用一台化00化aven Ze化PALS仪器),获得合成纳米载体尺寸(例如,直径)的测量值。 It will be transported by these synthetic nanocarriers suspended in a liquid (typically aqueous) medium, and dynamic light scattering (the DLS) (e.g., using a table of 00 of aven Ze of PALS instruments), to obtain a synthetic nanocarrier size (e.g. diameter) measurements. 例如,可W将具有合成纳米载体的悬浮液从水性缓冲剂中稀释到纯化水中W达到近似0.01至O.lmg/mL的最终的合成纳米载体悬浮液浓度。 For example, a final composite having a W nanosuspension concentration of the carrier synthetic nanocarriers from the aqueous suspension was diluted in buffer to purified water W reaches approximately 0.01 to O.lmg / mL of. 可W在一个适合的比色皿中直接制备稀释的悬浮液,或转移至该比色皿中W用于化S分析。 W may be a suitable cuvette diluted suspension prepared directly, or transferred to the cuvette of W S for analysis. 然后可W将该比色皿置于化S中,允许其平衡至受控的溫度,并且然后扫描充分的时间W在介质的粘度和样品的折射率的适当输入的基础上获得稳定且可再现的分布。 W can then be placed in the cuvette of S, allowing it to equilibrate to temperature controlled, and then the scan time sufficient to obtain a stable W based on appropriate input sample viscosity and refractive index of the medium and reproducible Distribution. 然后报告有效直径,或该分布的平均值。 Then reports the effective diameter, or the average of the distribution. 合成纳米载体的"尺寸"或"大小"或"直径"意指使用动态光散射获得的粒度分布的平均值。 Synthetic nanocarriers "size" or "size" or "diameter" means the average dynamic light scattering particle size distribution obtained.

[0112] "MHC"是指主要组织相容性复合体,即在大多数脊椎动物中发现的编码MHC分子的一个大的基因组区或基因家族,运些MHC分子将被加工的蛋白质的片段或表位展示在细胞表面上。 [0112] "MHC" refers to a large fragment of a genomic region or gene family major histocompatibility complex, i.e., found in most vertebrates encoding MHC molecules, MHC molecules to be transported some processed proteins or epitopes displayed on the cell surface. 细胞表面上的MHC:肤的呈递允许通过免疫细胞(通常是T细胞)来监视。 MHC on the cell surface: the skin allows monitored by presenting immune cells (typically a T-cell). 存在两大类MHC分子:I类和II类。 There are two broad classes of MHC molecules: Class I and Class II. 通常,I类MHC分子被发现在有核细胞上并且将肤呈递给细胞毒性T细胞。 Typically, class I MHC molecules are found on the skin of nucleated cells and presented to cytotoxic T cells. II类MHC分子被发现在某些免疫细胞(主要是巨隧细胞、B细胞W及树突状细胞)上,运些免疫细胞共同称为专职性APCdMHC区中最有名的基因是编码细胞表面上的抗原呈递蛋白的亚型。 Class II MHC molecules are found in certain immune cells (mainly tunnel giant cells, B cells and dendritic cells W), the transport of these immune cells collectively referred to as full-time most famous APCdMHC region gene encoding the cell surface antigen-presenting protein isoforms. 在人类中,运些基因被称为人白细胞抗原化LA)基因。 In humans, these genes are referred to transport human leukocyte antigens of LA) gene.

[0113] "非甲氧基封端的聚合物"意指具有W除了甲氧基之外的部分结束的至少一个末端的聚合物。 [0113] "Non-methoxy-terminated polymer" means W having at least one terminal end of the polymer portion other than a methoxy group. 在一些实施例中,该聚合物具有W除了甲氧基之外的部分结束的至少两个末端。 In some embodiments, the polymer having at least two terminal W of the portions other than the end of the methoxy group. 在其他实施例中,该聚合物不具有W甲氧基结束的末端。 In other embodiments, the polymer does not have a terminal end of the W methoxy. "非甲氧基封端的普朗尼克聚合物"意指除了在两个末端处具有甲氧基的线性普朗尼克聚合物之外的一种聚合物。 "Non-methoxy-terminated Pluronic polymer" means a linear addition polymer Pluronic having a methoxy group at both ends of a polymer. 如在此所提供的聚合物纳米颗粒可W包含非甲氧基封端的聚合物或非甲氧基封端的普朗尼克聚合物。 As provided herein comprise polymeric nanoparticles W Pluronic polymer may be non-methoxy-terminated polymer or methoxy terminated.

[0114] "获得"意指直接从一种材料中取得并且基本上不用改变和/或加工而使用。 [0114] "obtained" means directly made from one material and substantially without changing and / or processing used.

[0115] "药学上可接受的赋形剂"意指与列举的运些合成纳米载体一起使用W配制本发明的组合物的药理学上非活性的材料。 [0115] "Pharmaceutically acceptable excipient" W formulated material using pharmacologically inert compositions of the present invention means a synthetic nanocarriers transport these together with the exemplified. 药物学上可接受的赋形剂包含本领域中已知的各种材料,包括但不限于糖类(如葡萄糖、乳糖,等等)、防腐剂(如抗微生物剂)、复原助剂、着色剂、盐水(如憐酸盐缓冲盐水)、W及缓冲剂。 A pharmaceutically acceptable excipient known in the art comprises a variety of materials, including but not limited to, sugars (such as glucose, lactose, etc.), a preservative (antimicrobial agent), rehabilitation aids, colorants , saline (e.g. pity salt buffered saline), W, and buffers.

[0116] "治疗方案(Protocol)"是指针对受试者的一种或多种物质的任何给药方案。 [0116] "therapeutic regimen (Protocol)" means any dosing regimen against one or more species of the subject. 给药方案可W包括给药的量、频率和/或方式。 W dosing regimen can comprise administering an amount, frequency, and / or manner. 在一些实施例中,此种治疗方案可W用来向一个或多个测试受试者给予本发明的一种或多种组合物。 In some embodiments, W can be used for such a treatment regimen administration of one or more compositions of the present invention to one or more of the test subject. 然后可W对运些测试受试者中的免疫应答进行评定,W便确定该治疗方案是否对降低所不希望的免疫应答或产生所希望的免疫应答(例如,促进致耐受性作用)有效。 W can then be shipped to an immune response in the subject of these tests were evaluated, W will determine whether the treatment regimen to reduce undesirable immune responses or to generate a desired immune response (e.g., promoting a tolerogenic effect) effectively . 还可W代替或除了前面提到的运些免疫应答之外,对任何其他治疗性和/或预防性作用进行评定。 W may be transported in place of or in addition to those previously mentioned an immune response, any other therapeutic and / or prophylactic effect to be assessed. 可W使用在此提供的任何方法或本领域中已知的其他方法确定一个治疗方案是否具有所希望的作用。 W may be used any of the methods provided herein or other methods known in the art to determine whether a treatment regimen desired effect. 例如,可W从已经根据一个具体的治疗方案而被给予了在此提供的一种组合物的受试者中获得一群细胞,W便确定特异性免疫细胞、细胞因子、抗体等等是否被减少、产生、激活等等。 For example, W can be had from a particular treatment regimen in accordance with the subject was given a composition provided herein is a population of cells obtained, it is determined whether W specific immune cells, cytokines, antibodies and the like are reduced produce, activate, and so on. 用于检测免疫细胞的存在和/ 或数目的有用方法包括但不限于流式细胞检测方法(例如,FACS)和免疫组织化学方法。 The presence and / or for detecting the number of immune cells include, but are useful in a method (e.g., FACS) and immunohistochemistry method is not limited to flow cytometry. 用于免疫细胞标记的特异性染色的抗体和其他结合剂是商业上可获得的。 Antibodies and other binding agents specific immunocytochemistry for markers are commercially available. 此类试剂盒典型地包括用于多种抗原的染色试剂,运些染色试剂允许来自异质细胞群体的所希望的细胞群的基于FACS的检测、分离和/或定量。 Such kits typically include a plurality of antigens staining reagents, staining reagents allowing some transport FACS-based detection, isolation and / or quantitatively cell population from a heterogeneous cell population desired.

[0117] "提供受试者"是引起临床医师与受试者接触并且向其给予在此提供的一种组合物或在其上进行在此提供的一种方法的任何行为或一系列行为。 [0117] "Providing subject" is caused by contact with the subject and the clinician administering thereto a composition provided herein, or any acts of a method provided herein or series of acts thereon. 优选地,该受试者是对如在此所提供的一种致耐受性免疫应答有需要者。 Preferably, the subject is one kind of immune tolerance induced as provided herein answer those in need. 该行为或一系列行为可W是本身直接地或间接地,如但不限于,通过依赖某人的语言或行为采取行动的不相关的第Ξ方。 The act or series of acts may be W is itself directly or indirectly, such as, but not limited to, the action by relying on one's language or behavior of the Ξ party irrelevant.

[0118] "受试者"意指动物,包括溫血哺乳动物,如人类和灵长类;鸟类;家庭饲养动物或农场动物,如猫、狗、绵羊、山羊、牛、马W及猪;实验动物,如小鼠、大鼠W及豚鼠;鱼类;爬行动物;动物园和野生动物;等等。 [01] A "subject" refers to animals, including warm-blooded mammals, such as humans and primates; bird; household animals or farm animals, such as cats, dogs, sheep, goats, cattle, horses and swine W ; laboratory animals, such as mice, rats and guinea pigs W; fish; reptiles; zoo and wild animals; and the like.

[0119] "基本上无B细胞表位"是指不存在处于刺激B细胞应答的基本上激活的量的B细胞表位(在抗原的背景下是其本身、与一种载体结合、或与一种发明的组合物结合)。 [0119] "substantially no B-cell epitope" means substantially in the absence of stimulation of activated B cell response in an amount of B-cell epitopes (which is itself in the context of an antigen, in combination with a carrier, or binding composition of the invention). 在多个实施例中,基本上无B细胞表位的一种组合物不含可测量的量的抗原的B细胞表位。 In various embodiments, the B cell epitope substantially no B cell epitope composition does not contain a measurable amount of the antigen. 在其他实施例中,此种组合物可W包含可测量的量的抗原的B细胞表位,但所述量并不有效于产生一种可测量的B细胞免疫应答(在抗原的背景下是其本身、与一种载体结合、或与一种发明的组合物结合),如抗原特异性抗体产生或抗原特异性B细胞增殖和/或活性,或并不有效于产生一种显著可测量的B细胞免疫应答(在抗原的背景下是其本身、与一种载体结合、或与一种发明的组合物结合)。 In other embodiments, such compositions may comprise B cell epitope W measurable amounts of antigen, but the amount is not effective to produce an immune response B-cells one measurable (in the context of the antigen itself, in combination with a carrier, or in combination with a composition of the invention), such as antigen-specific antibodies or antigen-specific B cell proliferation and / or activity, or are not effective to produce a significant measurable B cell immune response (which is itself bound to a carrier in the context of an antigen, or in combination with an inventive composition). 在一些实施例中,一种显著可测量的B细胞免疫应答是在受试者中产生或将预期会产生不利的临床结果的B细胞免疫应答。 A significant B cell immune response is to produce a measurable or would be expected to have adverse clinical outcomes in a subject In some embodiments, a B cell immune response. 在其他实施例中,一种显著可测量的B细胞免疫应答是大于由对照抗原(例如,已知不包含该抗原的B细胞表位或不刺激B细胞免疫应答的抗原)产生的相同类型免疫应答(例如,抗原特异性抗体产生或抗原特异性B细胞增殖和/或活性)的水平的B细胞免疫应答。 In other embodiments, B is a cell of the immune response is measurable significantly greater than the same type of immune generated by control antigens (e.g., known not to contain B cell epitopes of the antigen or antigen-stimulated B cell immune response) response (e.g., antigen-specific antibodies or antigen-specific B cell proliferation and / or activity) levels of B cell immune response. 在一些实施例中,一种显著可测量的B细胞免疫应答(如抗体滴度的测量值(例如,通过化ISA))比由一种对照物(例如,对照抗原)产生的相同类型应答大2倍、3倍、4倍、5倍、6倍、7倍、8倍、9倍、10倍、15倍、20倍或更多倍。 In some embodiments, a significantly measurable B cell immune responses (as measured value (e.g., by chemical ISA) antibody titer) is larger than the same type of response generated from a control (e.g., control antigen) 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold or more. 在其他实施例中,基本上无B细胞表位的一种组合物是产生很少至不产生抗原特异性抗体滴度的组合物(在抗原的背景下是其本身、与一种载体结合、或与一种发明的组合物结合)的组合物。 In other embodiments, substantially no B cell epitope is a composition producing little to no antigen-specific antibody titer composition (which is itself in the context of an antigen, in combination with a carrier, the composition of the invention with one or a combination) of the composition. 此类组合物包括产生的抗体滴度(如EC50值)为小于500、400、300、200、100、50、40、30、20或10的那些。 Antibody titer (e.g., EC50 value) Such compositions include those produced 500,400,300,200,100,50,40,30,20 or less than 10. 在一些实施例中,一种显著可测量的B细胞免疫应答是比由一种对照物产生的相同类型应答大10 %、25 %、50 %、100 %、2倍、3倍、4倍、5倍、6倍、7倍、8倍、9倍、10倍、15倍、20 倍或更多倍的B细胞数目或增殖的测量值。 In some embodiments, B is a cell of the immune response is a response significantly measurable 10% higher than the same type generated from a control, 25%, 50%, 100%, 2 fold, 3 fold, 4 fold, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 15 times, 20 times or more the number of measurements or B cell proliferation. 用于测量B细胞应答的其他方法是本领域普通技术人员已知的。 Other methods for measuring B cell response to those of ordinary skill in the art.

[0120] 在多个实施例中,为了确保一种组合物基本上不包含B细胞表位,对抗原进行选择,使得运些抗原不包含用于与如在此所提供的与合成纳米载体偶联的B细胞表位。 [0120] Example, in order to ensure that a composition substantially does not comprise a B cell epitope of an antigen selected such that transport of these antigens, do not contain the synthetic nanocarriers as herein provided in a plurality of embodiments even B-cell epitopes linked. 在其他实施例中,为了确保一种组合物基本上不包含抗原的B细胞表位,产生与该抗原偶联的合成纳米载体并且测试其B细胞免疫应答(例如,抗原特异性抗体产生、B细胞增殖和/或活性)。 In other embodiments, in order to ensure that a composition substantially does not comprise B cell epitopes of an antigen, generates synthetic nanocarriers conjugated to the antigen and to test its B cell immune responses (e.g., produce antigen-specific antibodies, B cell proliferation and / or activity). 然后可W选择展现出所希望的特性的多种组合物。 W can then select compositions exhibit various desirable characteristics.

[0121] "-种或多种合成纳米载体"意指在自然中未发现的、并且在大小上具有小于或等于5微米的至少一个尺寸的离散物体。 [0121] "- one or more synthetic nanocarriers" means not found in nature, and less than or equal to 5 microns in at least one dimension size of discrete objects. 白蛋白纳米颗粒通常被包括为合成纳米载体,然而在某些实施例中,运些合成纳米载体并不包含白蛋白纳米颗粒。 Albumin nanoparticles typically comprise synthetic nanocarriers, however, in some embodiments, some synthetic nanocarrier transport does not contain albumin nanoparticles. 在多个实施例中,发明的合成纳米载体不包含壳聚糖。 In various embodiments, synthetic nanocarriers invention does not comprise chitosan. 在其他实施例中,发明的合成纳米载体不是基于脂质的纳米颗粒。 In other embodiments, the invention is not synthetic nanocarriers lipid-based nanoparticles. 在另外多个实施例中,发明的合成纳米载体不包含憐脂。 In further various embodiments, synthetic nanocarriers pity invention does not contain fat.

[0122] -种合成纳米载体可W是但不限于W下各项中的一种或多种:基于脂质的纳米颗粒(在此又称为脂质纳米颗粒,即构成它们结构的大多数材料是脂质的纳米颗粒)、聚合物纳米颗粒、金属纳米颗粒、基于表面活性剂的乳液、树枝状化合物、己基球、纳米线、病毒样颗粒(即,主要由病毒结构蛋白构成、但不是感染性的或具有低感染性的颗粒)、基于肤或蛋白质的颗粒(在此又称为蛋白质颗粒,即构成它们结构的大多数材料是肤或蛋白质的颗粒) (如白蛋白纳米颗粒)、和/或使用纳米材料的组合而发展的纳米颗粒(如脂质-聚合物纳米颗粒)。 [0122] - synthetic nanocarriers W may be but is not limited to, one or more of the W of: a lipid-based nanoparticle (also referred to herein lipid nanoparticles, i.e. they constitute the majority of the structure material is a lipid nanoparticles), polymer nanoparticles, metal nanoparticles, emulsion based surfactants, dendrimers, hexyl ball, nanowires, virus-like particles (i.e., mainly composed of viral structural proteins, but not infectious or has a low infectious particle), the particle-based skin or protein (also referred to herein proteinaceous particles, i.e. they constitute the majority of the material is a particulate skin structure or protein) (such as albumin nanoparticles), and / or combinations of materials and the development of nano nanoparticles (lipid - polymer nanoparticles). 合成纳米载体可W具有各种不同的形状,包括但不局限于:球形、立方形、锥形、长方形、圆柱形、螺旋管形,等等。 Synthetic nanocarriers W can have a variety of shapes, including but not limited to: spherical, cubic, conical, rectangular, cylindrical, toroidal, and the like. 根据本发明的合成纳米载体包含一个或多个表面。 The synthetic nanocarriers of the present invention comprises one or more surfaces. 可W适合用于实践本发明的示例性的合成纳米载体包含:(1)披露在格列夫(Gref)等人的美国专利5, 543,158中的可生物降解的纳米颗粒、(2)萨尔兹曼(Sal tzman)等人的公开美国专利申请20060002852的聚合物纳米颗粒、(3)戴斯蒙(DeSimone)等人的公开美国专利申请20090028910的光刻构建(lithogra地ically constructed)的纳米颗粒、(4)冯•艾德里安(von An化ian)等人的W0 2009/051837的披露内容、或(5)披露在佩纳德斯。 W may be for example synthetic nanocarriers used in the practice of the present invention comprises: (1) disclosed in Gref (Gref) et al., U.S. Patent No. 5, biodegradable nanoparticles 543,158, (2) Salzman (Sal tzman) et al., U.S. Patent application Publication 20060002852 polymeric nanoparticles, and (3) Desmond (DeSimone,) et al., U.S. Patent application Publication 20090028910 photolithography construct (lithogra to ically constructed) of nanoparticles, disclosure (4) • Adrian von (von An of ian) and others W0 2009/051837 or (5) disclosed in Pena Fernandez. enades)等人的公开美国专利申请2008/0145441中的纳米颗粒、(6)披露在德洛斯里奥斯(de los Rios) 等人的公开美国专利申请20090226525中的蛋白质纳米颗粒、(7)披露在西贝尔(Sebbel)等人的公开美国专利申请20060222652中的病毒样颗粒、(8)披露在己赫曼(Bachmann)等人的公开美国专利申请20060251677中的与核酸偶联的病毒样颗粒、(9)披露在W0 2010047839 A1或W0 2009106999 A2中的病毒样颗粒、(10)披露在P ·保利赛利(P.Paolicelli)等人的"可W高效结合并递送病毒样颗粒的表面修饰的基于化GA的纳米颗粒(Surface-modified PLGA-based Nanoparticles that can Efficiently Associate and Deliver Virus- l;Lke Particles),'纳米医学(Nanomedicine),5(6) :843-853(2010)中的纳米沉淀的纳米颗粒、或(11)披露在美国公开号2002/0086049中的稠亡细胞、稠亡小体、或合成或半合成的模拟物。在多个实施例中,合成纳米载体可^具有大 enades) et al., disclosed in U.S. Patent Application 2008/0145441 nanoparticles, (6) disclosed in de los Rios (de los Rios) et al., U.S. Patent Application Publication protein nanoparticles 20090226525, (7) discloses in Siebel (Sebbel) et al., U.S. Patent Publication application 20060222652 virus-like particles (8) have disclosed in Herman (of Bachmann) et al., U.S. Patent application Publication coupled to the nucleic acid of the virus-like particle 20060251677, (9) the virus-like particles are disclosed in W0 2010047839 A1 or in W0 2009106999 A2, (10) incorporated in the disclosure of efficient protection P · Li Saili (P.Paolicelli) et al., "W and can deliver virus-like particles surface-modified nanoprecipitation 843-853 (2010) in: 'nanomedicine (nanomedicine), 5 (6); nanoparticles (Lke particles Surface-modified PLGA-based nanoparticles that can Efficiently Associate and Deliver Virus- l) of the GA-based nanoparticles, or (11) Publication No. 2002/0086049 disclosure of fused cell death in the United States, fused dead bodies, or a synthetic or semi-synthetic analog thereof. in various embodiments, synthetic nanocarriers can have a large ^ 1:1、1:1.2、1:1.5、1:2、1:3、1:5、1:7、 或大于1:10的长宽比。 1: 1, 1: 1.2, 1: 1.5, 1: 2,1: 3,1: 5,1: 7, or an aspect ratio greater than 1:10.

[0123] 具有等于或小于约100皿、优选地等于或小于100皿的最小尺寸的根据本发明的合成纳米载体不包含具有使补体活化的径基的表面,或可替代地包含主要由不是使补体活化的径基的部分组成的表面。 [0123] equal to or less than about 100 dish, preferably equal to or smaller than the minimum size of 100 cuvette does not contain surface has a complement activation path group based synthetic nanocarriers of the present invention, or may alternatively comprise mainly not make complement activation group consisting diameter portion surface. 在一个优选的实施例中,具有等于或小于约lOOnm、优选地等于或小于lOOnm的最小尺寸的根据本发明的合成纳米载体不包含基本上使补体活化的表面, 或可替代地包含主要由不是基本上使活化补体的部分组成的表面。 In a preferred embodiment, equal to or less than about lOOnm, preferably equal to or smaller than the minimum size of lOOnm does not contain substantially cause complement activation surface, or may alternatively comprise mainly of the synthetic nanocarriers of the present invention are not substantially the activated complement-part surface. 在一个更优选的实施例中,具有等于或小于约lOOnm、优选地等于或小于lOOnm的最小尺寸的根据本发明的合成纳米载体不包含使补体活化的表面,或可替代地包含主要由不使补体活化的部分组成的表面。 In a more preferred embodiment, equal to or less than about lOOnm, preferably equal to or smaller than the minimum size of lOOnm is not included so that complement activation surface, or alternatively comprising the synthetic nanocarriers of the present invention consists essentially without complement activation part surface. 在多个实施例中,合成纳米载体排除病毒样颗粒。 In various embodiments, synthetic nanocarriers exclude virus-like particles. 在多个实施例中,合成纳米载体可W 具有大于1:1、1:1.2、1:1.5、1:2、1:3、1:5、1:7、或大于1:10的长宽比。 In various embodiments, synthetic nanocarriers W may have greater than 1: 1, 1: 1.2, 1: 1.5, 1: 2,1: 3,1: 5,1: 7, 1:10, or greater than the length and width ratio.

[0124] 叮细胞抗原"意指CD8巧细胞抗原或CD8+细胞抗原。乂D4巧细胞抗原"意指由CD8+T 细胞识别并且触发CD8巧细胞中的免疫应答的任何抗原,例如经由结合到II类主要组织相容性复合体分子(MHC)上的抗原或其一部分的呈递而被CD8巧细胞上的T细胞受体特异性地识别的抗原/'CD8巧细胞抗原"意指由CD8巧细胞识别并且触发CD8巧细胞中的免疫应答的任何抗原,例如经由结合到I类主要组织相容性复合体分子(MHC)上的抗原或其一部分的呈递而被CD8巧细胞上的T细胞受体特异性地识别的抗原。在一些实施例中,作为T细胞抗原的抗原也是一种B细胞抗原。在其他实施例中,该Τ细胞抗原并不也是一种B细胞抗原。Τ细胞抗原通常是蛋白类或肤类。 [0124] bite cell antigen "means an antigen or a CD8 cell Qiao antigen CD8 + cells. Qe D4 Qiao cell antigen" is meant a CD8 + T cells recognize antigen and trigger any coincidence CD8 cell immune response, e.g., via binding to II presentation of the antigen or a portion thereof on the category of major histocompatibility complex molecules (MHC) and is specifically recognized by T cell receptors on CD8 cell antigen clever / 'CD8 clever cell antigen "is meant a CD8 cells clever any antigen recognition and triggering an immune response of CD8 cells Qiao, e.g., via binding to an antigen or a portion thereof present in the class I major histocompatibility complex molecules (MHC) is skillfully CD8 T cell receptors on cells specifically recognizes the antigen. in some embodiments, a T cell antigen is an antigen, a B cell antigen. in other embodiments, the Τ cell antigen, a B cell antigen is not .Τ cell antigens are usually proteins or skin type.

[012引"效应性Τ细胞"被公认是运样一个术语,即是指:执行Τ细胞功能的Τ细胞,并且包括具有细胞毒性活性的Τ细胞、Τ辅助细胞、分泌细胞因子并且活化和/或引导其他免疫细胞的Τ细胞,等等。 [012 cited "Τ effector cells" is recognized as a sample transport term, which means: performing Τ cell function Τ cells, and includes a Τ cell cytotoxic activity, Τ helper cells, and secretion of cytokines and activation / or other immune cells Τ guide cells, and the like. 效应性Τ细胞包括,例如,ThO、Th 1、Th2、Th9、Th 17、W及效应记忆Τ细胞,运些细胞都是CD4+的,W及CD8+细胞毒性T细胞。 Τ effector cells include, for example, ThO, Th 1, Th2, Th9, Th 17, W Τ and effector memory cells, these cells are CD4 + transport of, W, and CD8 + cytotoxic T cells.

[0126] "治疗性蛋白"是指可W向受试者给予并且具有一种治疗性作用的任何蛋白质或基于蛋白质的疗法。 [0126] "therapeutic protein" refers to administering to a subject W having a therapeutic effect and any protein or protein-based therapeutics. 此类疗法包括蛋白质替代和蛋白质补充疗法。 Such therapies include protein replacement therapy and protein supplement. 此类疗法还包括给予外源或外来的蛋白质、抗体疗法、W及细胞或基于细胞的疗法。 Such therapy further comprises administration of exogenous or foreign protein, antibody therapy, W, and cells or cell-based therapies. 治疗性蛋白包括酶、酶辅助因子、激素、凝血因子、细胞因子、生长因子、单克隆抗体、W及多克隆抗体。 Therapeutic proteins include enzymes, enzyme cofactors, hormones, clotting factors, cytokines, growth factors, monoclonal antibodies, W, and polyclonal antibodies. 在此在其他地方提供了其他治疗性蛋白的实例。 In this example it provides other therapeutic proteins in other places. 可W在细胞之中、之上或由细胞产生治疗性蛋白,并且可W从此类细胞中获得治疗性蛋白、或W此类细胞的形式给予运些治疗性蛋白。 W in the cell may be, on or therapeutic protein produced by a cell, and W can be obtained from therapeutic proteins such cells, these cells or form W dosed some therapeutic protein. 在多个实施例中, 在W下细胞之中、之上或由W下细胞产生该治疗性蛋白:哺乳动物细胞、昆虫细胞、酵母细胞、细菌细胞、植物细胞、转基因动物细胞、转基因植物细胞等等。 In various embodiments, among the cells, or on the therapeutic protein produced by a cell under W at W: mammalian cells, insect cells, yeast cells, bacterial cells, plant cells, a transgenic animal cell, transgenic plant cell and many more. 可W在病毒转化的细胞之中、之上或由病毒转化的细胞产生该治疗性蛋白。 W may be in virally transformed cells, or to produce on the therapeutic protein by the transformed cell virus. 还可W在自体细胞之中、之上或由自体细胞产生该治疗性蛋白,运些自体细胞已经被转染、转导或W另外的方式操纵W表达该治疗性蛋白。 W may also be autologous cells in, on or by the autologous cells to produce the therapeutic protein, transport of these autologous cells has been transfected, transduced or otherwise manipulated W W expression of the therapeutic protein. 可替代地,可W作为一种核酸或通过将一种核酸引入到病毒、VLP、脂质体等等中来给予该治疗性蛋白。 Alternatively, W may be introduced as a nucleic acid or virus, VLPs, liposomes and the like by a nucleic administered in the therapeutic protein. 可替代地,可W从此类形式中获得该治疗性蛋白并且作为该治疗性蛋白本身而给予。 Alternatively, W may be obtained from the therapeutic protein such as the forms and the administration of therapeutic protein itself. 因此,受试者包括已经接受、正接受或将要接受上述任何物质的任何受试者。 Thus, the subject has received comprises, we are receiving or about to receive any subject of any of the foregoing. 此类受试者包括已经接受、正接受或将要接受W下各项的受试者:基因治疗;已经被转染、转导或W另外的方式操纵W表达一种治疗性蛋白、多肤或肤的自体细胞;或表达一种治疗性蛋白、多肤或肤的细胞。 Such subjects include has received, we are receiving or about to receive the subject under W: Gene therapy; has been transfected, transduced or otherwise manipulated W W express a therapeutic protein, or multiple skin autologous skin cells; or express a therapeutic protein, a multi-cell skin or skin.

[0127] "治疗性蛋白抗原"意指与一种治疗性蛋白相关联的抗原,该治疗性蛋白可W是、 或该治疗性蛋白的一部分可W是被呈递W供免疫系统的细胞进行识别并且可W产生针对该治疗性蛋白的一种所不希望的免疫应答(例如,产生治疗性蛋白特异性的抗体)。 [0127] "antigen therapeutic protein" is meant a therapeutic protein and an antigen associated with the therapeutic protein may be W, W, or a portion of the therapeutic protein is W-presenting cells for the immune system to identify W and may generate an immune response against one of the therapeutic protein is undesirable (e.g., to produce antibodies specific therapeutic protein). 治疗性蛋白抗原通常包括蛋白质类、多肤类、肤类、脂蛋白类,或被包含在或表达在细胞之中、之上或由细胞表达。 Therapeutic protein antigens typically comprises proteins, multi-skin type, skin type, lipoproteins, or contained within the cell or expressed in, on or expressed by a cell.

[0128] "致耐受免疫应答"意指可W导致对一种抗原或表达此种抗原的一种细胞、组织、 器官等等特异的免疫抑制的任何免疫应答。 [0128] "tolerogenic immune response" refers to an antigen may result in W or expression of any such a cellular immune antigen, tissue, organ-specific immunosuppression and the like response. 此类免疫应答包括对该抗原或表达此抗原的细胞、组织、器官等等特异的所不希望的免疫应答的任何降低、延迟或抑制。 Such an immune response to the antigen includes any reduction in expression of this antigen, or a cell, tissue, organ and the like are not specific to a desired immune response, delayed or suppressed. 此类免疫应答还包括对该抗原或表达此抗原的细胞、组织、器官等等特异的所希望的免疫应答的任何刺激、 产生、诱导、促进或募集。 Such an immune response to the antigen further comprises or express any antigen stimulation of this cell, tissue, organ and the like specific to the desired immune response is generated, inducing, promoting or recruitment. 因此,致耐受性免疫应答包括不存在或降低针对抗原的所不希望的免疫应答,运可W通过抗原反应性细胞W及存在或促进抑制性细胞来介导。 Accordingly, tolerogenic immune response comprises the absence or reduced immune response against the undesired antigen, cell W W may be transported and the presence or promotion of inhibiting cell mediated by an antigen reactivity. 在此所提供的致耐受性免疫应答包括免疫耐受性。 In the immune tolerance induced responses provided herein include immune tolerance. "产生致耐受性免疫应答"意指产生对一种抗原或表达此抗原的细胞、组织、器官等等特异的任何上述免疫应答。 "Generate tolerogenic immune response" is meant any of the above immune produce an antigen or a cell expressing this antigen, tissue, organ-specific response and the like. 该致耐受性免疫应答可W是MHC I类限制性呈递和/或MHC II类限制性呈递和/或B细胞呈递和/或由CDld进行的呈递等等的结果。 The tolerogenic immune response may result W is a MHC class I restricted presentation and / or MHC II-restricted presentation and / or B cells present and / or the like performed by the presentation CDld.

[0129] 致耐受性免疫应答包括CD4巧细胞、CD8巧细胞或B细胞增殖和/或活性的任何降低、延迟或抑制。 [0129] tolerogenic immune responses including CD4 clever cells, CD8 cells or cell proliferation Qiao B / or decreased activity of any delay or inhibition. 致耐受性免疫应答还包括抗原特异性抗体产生的降低。 Tolerance induced immune response further comprising reducing antigen specific antibody production. 致耐受性免疫应答还可W包括导致调节性细胞的刺激、诱导、产生或募集的任何应答,运些调节性细胞如CD4+ 化eg细胞、CD8巧reg细胞、Breg细胞等等。 Any response tolerogenic immune response may also cause irritation W includes regulatory cells, inducing, generating, or raised, transport these regulatory cells such as CD4 + cells of eg, CD8 clever reg cells, cells, etc. Breg. 在一些实施例中,该致耐受性免疫应答是导致转变为一种调节性表型的致耐受性免疫应答,该调节性表型的特征在于产生、诱导、刺激或募集调节性细胞。 In some embodiments, the tolerogenic immune response leading to into tolerogenic immune response to a regulatory phenotype, the regulatory phenotype characterized by the production, induction, stimulation or recruitment of regulatory cells.

[0130] 致耐受性免疫应答还包括导致CD4+化eg细胞和/或CD8+化eg细胞的刺激、产生或募集的任何应答。 [0130] tolerogenic immune response further comprises causing eg of CD4 + cells / and or any response of CD8 + cells stimulated eg, created or recruited. CD4巧reg细胞可W表达转录因子化xP3并且抑制炎症应答和自身免疫性炎性疾病(自身免疫性疾病中的人调节性T细胞(Human regulatory T cells in autoimmune diseases),科夫塔诺维奇•化(Cve1:anovich 化)、哈弗《DA(化fler DA),免疫学当前观点(Curr Opin Immunol. )2010年12月;22(6) :753-60,调节性Τ细胞和自身免疫性(Regulatory Τ cells and autoimmunity),维拉*J(Vila J)、艾萨克*JD(Isaacs JD)、安德森*AE(Anderson AE),血液学当前观点(Curr Opin 胎matol.)2009年7月;16 (4) :274-9)。 Reg cells may be CD4 Qiao W xP3 of transcription factors and suppression of the inflammatory response and autoimmune inflammatory diseases (autoimmune diseases in human regulatory T cells (Human regulatory T cells in autoimmune diseases), Kofta Norwich • of (Cve1: anovich of), Harvard "DA (of fler DA), current Opinion in Immunology (Curr Opin Immunol.) 2010 Nian Dec; 22 (6): 753-60, regulatory Τ cells and autoimmune (Regulatory Τ cells and autoimmunity), Vera * J (Vila J), Isaac * JD (Isaacs JD), Anderson * AE (Anderson AE), hematology current July 2009 view (Curr Opin tire matol.) ; 16 (4): 274-9). 此类细胞还抑制Τ细胞对B细胞的帮助并且诱导对自身和外来抗原的耐受性(基于FOXP3+调节性T细胞活化和扩增的过敏和自身免疫性的治疗性方法(Therapeutic approaches to allergy and autoimmunity based on FoxP3+regulatory T-cell activation and expansion),宫良· M(Miyara M)、永《RXling K)、板口·(Sakaguchi S),过敏临床免疫学期刊(J Allergy Clin Immunol) ,2009年4月;123(4) :749-55)。 Such cells also inhibited Τ cells help B cells and induction of tolerance to self and foreign antigens (based FOXP3 + regulatory T cell activation and expansion of allergies and autoimmune therapeutic methods (Therapeutic approaches to allergy and autoimmunity based on FoxP3 + regulatory T-cell activation and expansion), Gongliang · M (Miyara M), Wing "RXling K), the plate opening · (Sakaguchi S), Journal of allergy clinical Immunology (J allergy Clin Immunol), 2009 April; 123 (4): 749-55). 当抗原由APC上的II类蛋白质呈递时,CD4+化eg细胞识别运种抗原。 When the antigen presented by class II proteins on APC, CD4 + cells recognize transport of eg antigens. 识别由I类(和化-1)呈递的抗原的CD8+Treg细胞还可W抑制T细胞对B细胞的帮助,并且导致诱导对自身和外来抗原二者的耐受性的抗原特异性抑制的活化。 Identifying presented by class I (and of -1) antigen CD8 + Treg cells can also suppress W T cells help B cells, and antigen results in the induction of tolerance to both self and foreign antigens specific inhibition of activation. 已经显示,Qa-1与CD8+化eg细胞的相互作用的破坏可使免疫应答失调,并且导致自身抗体形成和自身免疫性致命性系统性红斑狼疮的产生(金化im)等人,自然(化Uire),2010年9月16日,467(7313):328-32)。 It has been shown, Qa-1 interaction eg destruction of cells CD8 + immune response can disorders, and leads to formation of autoantibodies and autoimmune produce fatal systemic lupus erythematosus (alloying im) et al., Nature (of Uire), 2010 Nian 9 16, 467 (7313): 328-32). 还已经显示,〔08+化6邑细胞可抑制包括类风湿性关节炎和结肠炎的自身免疫性炎性疾病的模型(自身免疫性关节炎中的CD4 + CD25 +调节性T细胞(CD4 + CD25 + regulatory T cells in autoimmune arthritis),欧.S(0h S)、兰金*AL(Rankin AL)、卡顿*AJ(Caton AJ),免疫性评论(Immunol Rev) ,2010年1月;233(1) :97-111,炎性肠病中的调节性Τ细胞(Regulatory Τ cells in inflammatory bowel disease).博登· EK(Boden EK)、其if 奈普· SB (Snapper SB),肠胃病学当前观点(Curr Opin Gastroenterol),2008年11 月;24(6): 733-41)。 It has also been shown that model [6 08+ cells suppressed Yap including rheumatoid arthritis and inflammatory colitis, autoimmune diseases (CD4 + CD25 + autoimmune arthritis in regulatory T cells (CD4 + CD25 + regulatory T cells in autoimmune arthritis), Europe .S (0h S), Rankin * AL (Rankin AL), Caton * AJ (Caton AJ), autoimmune comments (Immunol Rev), 2010 Jan; 233 (1): 97-111, inflammatory bowel disease Τ regulatory cells (regulatory Τ cells in inflammatory bowel disease) Boden · EK (Boden EK), which if naproxen · SB (Snapper SB), gastroenterology current view (Curr Opin Gastroenterol), November 2008; 24 (6): 733-41). 在一些实施例中,提供的运些组合物可W有效地产生两种类型的应答(CD4巧reg和CD8+化eg)。 In some embodiments, these compositions provide a transport W may be effective to produce the two types of response (CD4 and CD8 + reg clever of eg). 在其他实施例中,可W在其他免疫细胞(如巨隧细胞、iNKT细胞等等)中诱导化xP3,并且在此提供的运些组合物同样可W产生运些应答中的一种或多种。 In other embodiments, W may be in other immune cells (e.g., tunneling giant cells, of iNKT cells, etc.) of XP3 induced, transported and provided these compositions likewise W of generating a response to one or more of these transport species.

[0131] 致耐受性的免疫应答还包括但不限于:诱导调节性细胞因子,如化eg细胞因子;诱导抑制性细胞因子;抑制炎性细胞因子(例如,比-4、IL-化、化-5、TNF-a、比-6、GM-CSF、WN- 丫、化-2、化-9、化-12、化-17、化-18、化-21、化-22、化-23、M-CSF、C反应性蛋白、急性期蛋白、趋化因子(例如,]\〇"-1、1?4肿65、]\〇口-化、]\〇口-10、]\〇6、口4(:或1口-1〇);产生抗炎性细胞因子(例如,化-4、化-13、化-10等等)、趋化因子(例如,C化-2、CXCL8)、蛋白酶(例如,MMP-3、 MMP-9)、白Ξ締(例如,切sLT-1、切sLT-2)、前列腺素(例如,PGE2)或组胺;抑制向Th 17、化1 或化2免疫应答的极化;抑制效应细胞特异性的细胞因子:化17(例如,化-17、化-25)、化1 (IFN-丫)、Th2(例如,比-4、化-13);抑制Thl特异性、化2特异性或TH17特异性的转录因子; 抑制效应T细胞的增殖;诱导效应T细胞的调亡;诱导致 [0131] tolerogenic immune response further include, without limitation: induction of regulatory cytokines, such as cytokines, eg of; cytokines induced inhibition; inhibition of inflammatory cytokines (e.g., ratio of -4, IL- of, of -5, TNF-a, than -6, GM-CSF, WN- Ah, of -2, -9 oriented, of 12, of -17, -18 of, of -21, -22 of, of - 23, M-CSF, C-reactive protein, acute phase proteins, chemokines (e.g.,] \ square "swelling 65 4-1,1,] \ square mouth -? technology,] \ -10 port square,] \ 〇6, port 4 (: or a -1〇); produce anti-inflammatory cytokines (e.g., chemical -4, of -13, -10 and the like of), chemokines (e.g., C of -2, of CXCL8), proteases (e.g., MMP-3, MMP-9), white Ξ association (e.g., cut sLT-1, cut sLT-2), prostaglandins (e.g., of PGE2) or histamine; suppressed Th 17, of 1 or 2 of the polarization of the immune response; effector cell specific inhibition of cytokines: Chemical Formula 17 (e.g., of -17, -25 of), of 1 (the IFN-Ah), Th2 (e.g., ratio of -4, of -13); Thl-specific inhibition, specificity for 2 or TH17-specific transcription factor; inhibit the proliferation of effector T cells; the induction of effector T cell apoptosis; induced lead 受性的树突状细胞特异性的基因; 诱导化xP3表达;抑制I姐诱导或I巧介导的免疫应答;抑制抗体应答(例如,抗原特异性抗体的产生);抑制T辅助细胞应答;产生TGF-β和/或IL-10;抑制自身抗体的效应子功能(例如, 细胞耗尽、细胞或组织损伤、或补体活化的抑制);等等。 Of specific genes by dendritic cells; xP3 induction of expression; inhibition induced sister I or I-mediated immune responses clever; inhibition of antibody responses (e.g., produce antigen-specific antibodies); inhibition of T helper cell response; produce TGF-β and / or IL-10; autoantibody suppression effector function (e.g., cell depletion, cell or tissue injury, complement activation or inhibition); and the like.

[0132] 任何上述的免疫应答可W在一个或多个动物模型中体内测量或可W在体外测量。 [0132] Any of the above W may be measured in vivo immune response in one or more animal models or W may be measured in vitro. 本领域普通技术人员熟悉此类体内或体外测量。 Those of ordinary skill in the art is familiar with such in vitro or in vivo measurements. 所不希望的免疫应答或致耐受性免疫应答可W使用例如评定免疫细胞数目和/或功能的方法、四聚物分析、ELISP0T、基于流式细胞检测的细胞因子表达、细胞因子分泌的分析、细胞因子表达谱绘制、基因表达谱绘制、蛋白质表达谱绘制、细胞表面标记分析、基于PCR的免疫细胞受体基因用法的检测(参见T ·克雷(T.Clay)等人/'用于监测针对癌症的主动免疫治疗的细胞免疫应答的测定(Assays for Monitoring Cellular Immune Response to Active Immunotherapy of Cancer)"临床癌症研究(Clinical Cancer Research)?: 1127-1135(2001))等等来监测。所不希望的免疫应答或致耐受性免疫应答还可W使用例如评定血浆或血清中的蛋白质水平的方法、免疫细胞增殖和/或功能测定等等来监测。在一些实施例中,致耐受性免疫应答可W通过评定化xP3 的诱导来监测。另外,在实例中更详细地描述了具体方法。 The method of undesired immune response or tolerogenic immune response can be assessed for example, the number W of immune cells and / or function, tetramer analysis, ELISP0T, based on flow cytometry cytokine expression, and analysis of cytokine secretion cytokine expression profile drawing, drawing gene expression profiling, proteomic expression drawing, cell surface marker analysis, based on the detection of immune cell receptor gene usage in the PCR (see Cray T · (T.Clay) et / 'for monitoring assay (Assays for monitoring cellular immune response to active immunotherapy of cancer) "clinical cancer Research (clinical cancer Research) ?: 1127-1135 (2001)) and the like immune response to active immunotherapy against cancer cells was monitored. the the method of unwanted immune responses or tolerogenic immune response may be assessed using, for example W or plasma protein levels of the serum, the immune cell proliferation and / or functional assay to monitor the like. in some embodiments, the tolerogenic W immune response can be monitored by evaluation of the induced xP3. Further, the specific methods described in the examples in more detail.

[0133] 优选地,致耐受性免疫应答导致对在此描述的疾病、素乱或病症的发展、进展或病理的抑制。 [0133] Preferably, the tolerogenic immune response results in a disease described herein, the development of hormone disorder or condition, or inhibiting the progression of pathology. 可W用具有此类疾病、素乱或病症的动物模型来测量运些发明的组合物是否可W导致对在此描述的疾病、素乱或病症的发展、进展或病理的抑制。 W can having such a disease, disorder or condition prime animal models used to measure whether the operation of the invention these compositions can result in W described herein disease, disorder or condition developing prime, or inhibiting the progression of pathology. 在一些实施例中,可W 通过确定临床终点、临床功效、临床症状、疾病生物标记和/或临床得分来评定所不希望的免疫应答的降低或致耐受性免疫应答的产生。 In some embodiments, W may be determined by clinical endpoint, clinical efficacy, the clinical symptoms and biological markers of disease and / or clinical score to assess reduction or tolerance induced immune response undesired immune responses. 还可W用诊断试验评定所不希望的免疫应答或致耐受性免疫应答,W便评定在此所提供的疾病、素乱或病症的存在或不存在。 W may also be assessed by immunization with undesirable diagnostic test response or tolerogenic immune response, W will be assessed provided herein disease, disorder or condition prime presence or absence. 可W进一步通过测量受试者中的治疗性蛋白水平和/或功能的方法来评定所不希望的免疫应答。 W may further be assessed undesired immune response by a method of measuring protein levels in a subject a therapeutic and / or functions. 在多个实施例中,用于监测或评定所不希望的过敏性应答的方法包括通过皮肤反应性和/或过敏原特异性的抗体产生评定受试者中的一种过敏性应答。 In various embodiments, a method for allergic response to monitoring or assessing include undesirable reactions and / or allergen-specific antibody producing an allergic response in a subject by assessing the skin.

[0134] 在一些实施例中,可W在给予在此提供的合成纳米载体的一种组合物之前和/或在给予一种可移植的移植物或治疗性蛋白或暴露于一种过敏原之前监测或评定受试者中的一种所不希望的免疫应答或一种致耐受性免疫应答的产生。 Before [0134] In some embodiments, W can be administered in a composition of synthetic nanocarriers provided herein and / or administration of a transplantable graft or therapeutic protein or exposure to an allergen before the original the monitoring or assessing a subject produce an unwanted immune response or one tolerogenic immune response. 在其他实施例中,可W在给予在此提供的合成纳米载体的一种组合物之后和/或在给予一种可移植的移植物或治疗性蛋白或暴露于一种过敏原之后评定一种所不希望的免疫应答或致耐受性免疫应答的产生。 After the other embodiments, W can be administered in a composition of synthetic nanocarriers provided herein and / or administration of a transplantable graft or therapeutic protein or after exposure to an allergen one kind of assessment generating undesirable immune responses or tolerogenic immune response. 在一些实施例中,在给予合成纳米载体的该组合物之后、但在给予一种可移植的移植物或治疗性蛋白或暴露于一种过敏原之前完成该评定。 In some embodiments, following administration of the synthetic nanocarriers compositions, but administration of a transplantable graft or therapeutic protein or before exposure to an allergen to complete the evaluation. 在其他实施例中,在给予一种可移植的移植物或治疗性蛋白或暴露于一种过敏原之后、但在给予该组合物之前完成该评定。 In other embodiments, the administration of a transplantable graft or therapeutic protein or after exposure to an allergen, but the composition is administered prior to the completion of the evaluation. 在再其他的实施例中,在给予运些合成纳米载体和给予一种可移植的移植物或治疗性蛋白或暴露于一种过敏原二者之前进行该评定,而在又其他的实施例中,在给予合成纳米载体和给予一种可移植的移植物或治疗性蛋白或暴露于一种过敏原二者之后进行该评定。 In still other embodiments, the dosed administration of these synthetic nanocarriers and a portable graft or therapeutic protein or both prior to exposure to an allergen for the original evaluation, and in still other embodiments , synthetic nanocarriers administration and administration of a portable therapeutic protein or grafts, or after exposure to an allergen for both the original assessment. 在另外的多个实施例中,在给予运些合成纳米载体和/或给予一种可移植的移植物或治疗性蛋白或暴露于一种过敏原之前和之后都进行该评定。 In further various embodiments, the administration of some synthetic nanocarriers transport and / or administration of one or transplantable graft therapeutic protein or exposure to an allergen for both before and after the evaluation. 在再其他的实施例中,在受试者上进行多于一次的该评定,W便确定令人希望的免疫状态被维持在该受试者中,该受试者如患上或正处于患上一种炎性疾病、一种自身免疫疾病、一种过敏症、器官或组织排斥或移植物抗宿主病的风险中的受试者。 In still other embodiments, the evaluation performed more than once in the subject, W will be desirable to determine the immune status is maintained in the subject, the subject is at risk or is suffering from an on an inflammatory disease, an autoimmune disease, an allergic disease, organ or tissue rejection or graft-versus-host disease risk in subjects. 其他受试者包括已经经历或将要经历移植的那些受试者W及已经接受、正在接受或将要接受一种治疗性蛋白的那些受试者,针对该治疗性蛋白,运些受试者已经经历、正在经历或预期会经历一种所不希望的免疫应答。 Other subjects include already undergone or will undergo transplant W and those subjects have received, are receiving or about to receive those subjects a therapeutic protein for which a therapeutic protein, shipped some subject has undergone , immunity is undergoing or expected to undergo one undesirable response.

[0135] 可W通过测定一个或多个抗体滴度来评定抗体应答。 [0135] W may be determined by one or more antibody titer antibody response was assessed. "抗体滴度"意指抗体产生的可测量水平。 "Antibody titer" means a measurable levels of antibody production. 用于测量抗体滴度的方法是本领域中已知的并且包括酶联免疫吸附测定化LISA)。 A method for measuring antibody titer are known in the art and include enzyme linked immunosorbent assays of LISA). 在多个实施例中,该抗体应答可W被定量为例如抗体的数目、抗体的浓度、或滴度。 In various embodiments, the antibody response may be quantified, for example, the number W of an antibody, antibody concentration or titer. 运些值可W是绝对的或它们可W是相对的。 These values ​​can be transported W are absolute or they may be W is relative. 用于定量抗体应答的测定包括抗体捕获测定、酶联免疫吸附测定化LISA)、抑制液相吸附测定(inhibition liquid phase absolution assay,ILPAA)、火箭免疫电泳(rocket Immunoelectrophoresis,RIE)测定、W 及线状免疫电泳(line immunoelect;rophoresis,LIE)测定。 For the quantitative determination of the antibody response include antibody capture assays, enzyme-linked immunosorbent assay of LISA), a liquid phase inhibition assay (inhibition liquid phase absolution assay, ILPAA), rocket immunoelectrophoresis (rocket Immunoelectrophoresis, RIE) determination, W, and the line shaped immunoelectrophoresis (line immunoelect; rophoresis, LIE) was measured. 当比较一种抗体应答与另一种抗体应答时,优选地使用相同类型的定量值(例如,滴度)和测量方法(例如,ELISA)进行该比较。 When compared with another one antibody response antibody response, preferably using a quantitative value (e.g., titer) and measurement methods (e.g., ELISA) performed the same type of comparison.

[0136] 用于测量抗体滴度的化ISA方法(例如,一种典型的夹屯、化ISA)可W由W下步骤组成:(i)制备一种化ISA-板包被材料,使得感兴趣的抗体祀标与一种底物聚合物或其他适合的材料偶联;(ii)在一种水溶液(如PBS)中制备该包被材料,并且将该包被材料溶液递送至一个多孔板的孔中用于使该包被层过夜沉积在该多孔板上;(iii)用洗涂缓冲液(如含0.05%吐溫-20的PBS)彻底洗涂该多孔板W去除过量的包被材料;(iv)通过施加一种稀释剂溶液(如含10%胎牛血清的PBS)来封闭该板的非特异性结合;(V)用洗涂缓冲液将该封闭/稀释剂溶液从该板上洗涂下来;(Vi)根据需要用稀释剂对含有抗体和适当标准品(阳性对照)的一种或多种血清样品进行稀释,W获得适当地使ELISA应答饱和的浓度;(vii)在该多孔板上对运些血浆样品进行连续稀释,运样W涵盖适用于产生化ISA应答曲线 [0136] The method for measuring the antibody titer of the ISA (e.g., a typical clamp Tun, of ISA) may be W W by the steps of: (i) preparing a coating material of ISA- plate, so that the sense interest antibody labeled with one Si substrate polymer conjugation or other suitable material; the coating material (ii) is prepared in an aqueous solution (e.g., PBS), and the coating material was delivered to a perforated plate the hole for the coating layer is deposited on the porous plate overnight; (iii) washcoated with buffer (e.g. containing PBS 0.05% Tween-20) of the porous plate coated thoroughly washed to remove excess coating W material; (iv) to block nonspecific binding of the board by applying a diluent solution (e.g., containing 10% fetal bovine serum PBS); (V) washcoated with the blocking buffer / diluent solution from the plate wash down the coating; (Vi) to one or more diluted serum sample containing the antibody and appropriate standard (positive control) with the required diluent, W ELISA suitably obtained that the response saturation concentration; (VII) in the porous plate op some plasma samples serially diluted sample transport W cover adapted to generate a response curve of ISA 的浓度范围;(viii)解育该板W提供抗体-祀标结合;(ix)用洗涂缓冲液洗涂该板W去除未结合抗原的抗体;(X)添加适当浓度的处于相同稀释剂中的一种第二检测抗体,如能够结合第一抗体的一种偶联生物素的检测抗体;(xi)用所施加的该检测抗体解育该板,随后用洗涂缓冲液进行洗涂;(xii)添加将与生物素化的抗体上发现的生物素结合的酶(如链霉亲和素-HRP (辣根过氧化物酶))并且进行解育;(xiii)洗涂该多孔板;(xiv)将一种或多种底物(如TMB 溶液)添加至该板上;(XV)当显色完全时施加一种终止溶液(如2N硫酸);(xvi)在针对该底物的特定波长下读取运些板孔的光密度(450nm减去570nm处的读数);(xvi)对数据应用一个适合的多参数曲线拟合,并且将半最大有效浓度巧巧0)定义为达到运些板标准品的半最大值0D值时的曲线上的浓度。 Concentration range; (VIII) W of the plate to provide sterile solutions of antibody - labeled binding worship; (IX) washcoated with wash buffer W of the plate coated remove unbound antibody antigen; (X-) addition of a suitable diluent at the same concentration the one second detection antibody, such as an antibody capable of binding to the detection of a first antibody conjugated to biotin; (XI) with the detection antibody incubation solution applied to the plate, followed by washing with a wash coat coating buffer ; (XII) was added to the enzyme (e.g., streptavidin, avidin-HRP (horseradish peroxidase)) and incubated for de-biotin binding with an antibody found biotinylated; (XIII) wash-coated porous plate; (XIV) one or more substrates (e.g., TMB solution) was added to the plate; (XV) when applying one color when fully stop solution (e.g. 2N sulfuric acid); (XVI) in the bottom for the optical density (450nm minus the reading at 570nm) of these read operation at a specific wavelength plate hole thereof; (XVI) applied to the data for a multi-parameter curve fitting, and the half-maximal effective concentration Qiaoqiao 0) is defined is a concentration on the curve at half maximum of these transport 0D value of the standard plate.

[0137] "可移植的移植物"是指可W给予一位受试者的生物材料,如细胞、组织W及器官(全部或部分地)。 [0137] "transplantable graft" refers to a biological material W may be administered to a subject, such as cells, tissues and organs, W (fully or partially). 可移植的移植物可W是例如一种生物材料(如器官、组织、皮肤、骨骼、神经、腫、神经元、血管、脂肪、角膜、多能细胞、分化细胞(在体内或体外获得或衍生的))等的自身移植物、同种异体移植物、或异种移植物。 Portable grafts may be, for example, W is a biological material (such as organ, tissue, skin, bone, nerve, swelling, neurons, blood vessels, fat, cornea, pluripotent cells, differentiated cells (obtained or derived in vivo or in vitro a)) and the like autograft, allograft, or xenograft. 在一些实施例中,可移植的移植物例如由软骨、骨骼、细胞外基质、或胶原基质形成。 In some embodiments, the graft may be formed from, for example, grafts of cartilage, bone, extracellular matrix, or collagen matrices. 可移植的移植物还可W是单细胞、细胞悬浮液W及可W被移植的组织和器官中的细胞。 Portable graft W may also be a single cell, cell suspensions W and W can be transplanted cells in tissues and organs. 可移植的细胞典型地具有一种治疗功能,例如在受体受试者中缺乏或减退的功能。 Transplantable cells typically have a therapeutic function, such as lack of or decreased function in a recipient subject. 可移植的细胞的一些非限制性实例是β细胞、肝细胞、造血干细胞、神经干细胞、神经元、胶质细胞、或成髓銷细胞。 Some non-limiting examples of transplantable cells is β cells, liver cells, hematopoietic stem cells, neural stem cells, neurons, glial cells, or into cells marrow pin. 可移植的细胞可W是未修饰的细胞, 例如从供体受试者获得的并且可用于移植而没有任何遗传或表观遗传修饰的细胞。 W transplantable cells may be unmodified cells may be used for transplantation, for example, and without any genetic or epigenetic modification of cells obtained from a donor subject. 在其他实施例中,可移植的细胞可W是被修饰的细胞,例如获得自具有遗传缺陷、其中该遗传缺陷已得到纠正的受试者的细胞;或来源于重新编程的细胞的细胞,例如来源于获得自受试者的细胞的分化细胞。 In other embodiments, W can be transplanted cells may be modified cells, for example, obtained from a genetic defect, which has been corrected the genetic defect in cells of a subject; reprogramming or from cells, e.g. obtaining differentiated cells derived from the subject cell.

[0138] "移植"是指将一个可移植的移植物(例如,从供体受试者、从体外来源(例如分化的自体或异体原生的或诱导的多能细胞))转移(移动)到一个受体受试者中和/或在同一受试者中从一个身体部位转移(移动巧Ij另一个身体部位的过程。 [0138] "transplant" refers to a portable grafts (e.g., from a donor subject, from outside sources (e.g., differentiated autologous or allogeneic native or induced pluripotent cells)) transferred (moved) to a recipient subject and / or metastasis in the same subject (Ij Qiao another body part is moved from one body part.

[0139] "所不希望的免疫应答"是指由暴露于抗原而引起的、促进或加剧在此提供的疾病、素乱或病症(或其症状)的、或是在此提供的疾病、素乱或病症的症状的任何所不希望的免疫应答。 [0139] "undesirable immune response" refers to exposure to an antigen and cause, promote, or exacerbate disease provided herein, prime disorder or condition (or symptoms thereof), or diseases provided herein, prime any undesired immune disorder symptoms or condition response. 此类免疫应答通常对受试者的健康具有负面影响或表明受试者的健康的负面影响。 Such immune responses typically have a negative impact on healthy subjects or indicate negative health effects of the subject. 所不希望的免疫应答包括抗原特异性的效应性T细胞(例如CD4巧细胞、CD8巧细胞)增殖和/或活性。 Undesired immune responses include antigen-specific effector T cells (e.g. CD4 Qiao cells, CD8 cells Qiao) proliferation and / or activity. 因此,所希望的免疫应答包括效应性T细胞的缺失、抑制运类细胞的刺激或活化、抑制效应性T细胞增殖、抑制由效应性T细胞产生细胞因子,等等。 Thus, the desired immune response comprises deletion of effector T cells, stimulation or inhibition of the activation of cell-based transport, inhibition of proliferation of effector T cells, inhibit the production of cytokines by the effector T cells, and the like. 当CD4巧细胞活性受到抑制时,运种抑制还可W导致B细胞免疫应答的降低。 When skillfully CD4 cell activity is inhibited, inhibition of seed transport W may lead to reduced B cell immune response. 多种用于测试运些免疫应答的方法在此被提供、或另外是本领域普通技术人员已知的。 These various test run for an immune response are provided herein, or in addition to those of ordinary skill in the art.

[0140] C.发明组合物 [0140] C. The composition of the invention

[0141] 在此提供了致耐受性方法和有关组合物,运些方法包括给予合成纳米载体组合物,运些合成纳米载体组合物包含免疫抑制剂和一种抗原的Μ肥I类限制性表位和/或MHC II类限制性表位。 [0141] In this method provides tolerance induced and related compositions, comprising administering to transport these synthetic nanocarriers compositions, shipped some class I-restricted Μ fertilizer composition comprising a synthetic nanocarrier immunosuppressants and one antigen epitopes and / or MHC class II restricted epitopes. 此类方法和组合物有用于降低效应性Τ细胞数目和/或功能并促进对包含运些表位的抗原特异的致耐受性免疫应答的产生。 Such methods and compositions for reducing the number of generated have Τ effector cells and / or function and to promote immune tolerance induced by antigen specific operation comprising these epitopes response. 可W向希望有致耐受性免疫应答的受试者给予运些组合物。 W may be desired to have induced an immune response in the subject resistant dosed these compositions. 此类受试者包括已经患上或正处于患上一种炎性疾病、一种自身免疫性疾病、一种过敏症、器官或组织排斥、或移植物抗宿主病的风险中的那些受试者。 Such subjects include risk of suffering or already suffering from an inflammatory disease is in, an autoimmune disease, an allergic disease, organ or tissue rejection, or graft versus host disease in those test By. 此类受试者还包括已经被、正在被或将要被给予一种治疗性蛋白的那些受试者,该受试者已经经历或预期会经历一种针对该治疗性蛋白的所不希望的免疫应答。 Such subjects include also has been, is being or will be given to those subjects a therapeutic protein, the subject has undergone or will undergo one expected that the therapeutic protein against unwanted immune response. 此类受试者还包括已经经历或将要经历移植的那些受试者。 Such subjects also include those subject has undergone or will undergo transplantation.

[0142] 优选地,本发明的组合物导致效应性Τ细胞的缺失和/或其活性的降低。 [0142] Preferably, the compositions of the present invention results in a reduction absence of effector cells Τ and / or activity. 效应性Τ细胞的表面标记表达谱和细胞因子分泌概况是本领域已知的(例如,如在拉格(Rag),免疫学: 一本教科书,阿尔法科技国际有限公司(Immunology:a Textbook,Alpha Science Inti Ltd) (2005年8月15日),ISBN-10:1842652559(例如第13章)中所描述的)并且可W用来评定所提供的运些组合物的作用。 Expression profile of surface markers and cytokine secretion profiles Τ effector cells are known in the art (e.g., as in La (Rag), Immunology: a textbook, Alpha Technology International Limited (Immunology: a Textbook, Alpha Science Inti Ltd) (15 August 2005), ISBN-10: 1842652559 (e.g. chapter 13) as described) and W may be used to assess the effect provided by these compositions is transported. 效应性T细胞的示例性群体包括所有促进、起始、增强或支持例如通过CD4+CD25-细胞和CD8+CD45RA+细胞、W及还有记忆细胞(CD4+CD25+FOXP3-)引起的免疫应答的那些。 Exemplary effector T cell population includes all promotion, starting, e.g., by enhancing or supporting CD4 + CD25- cells and CD8 + CD45RA + cells, W, and also memory cells (CD4 + CD25 + FOXP3-) induced an immune response Those ones. 可W用于鉴定效应性T细胞的其他表面标记是本领域技术人员已知的, 并且运类标记包括但不限于化-12受体(α和β1链)W及细胞因子受体CXCR3和CCR5(Thl细胞);化-1受体,ST2L/T1 (比-1RI样分子),趋化因子受体CXCR4、CCR3、CCR4、CCR7和CCR8、W 及CD30(Th2细胞)。 W may be used to identify additional surface markers effector T cells are known to the skilled person, and include but are not limited transport class labels of -12 to receptor (β1 chain and [alpha]) W and cytokine receptors CXCR3 and CCR5 (of Thl cells); of-1 receptor, ST2L / T1 (ratio -1RI like molecules), chemokine receptor CXCR4, CCR3, CCR4, CCR7 and CCR8, W and CD30 (Th2 cells). 效应性T细胞进一步分泌细胞因子,例如IFN-丫、IL-2和/或TNF-f3(Thl细胞);比-4、比-5、比-13(Th2细胞)、化-9(化9)、比-17(Thl7)、化-22(Th22),或其他炎性细胞因子。 Effector T cells secrete cytokines further, e.g. Ah IFN-, IL-2 and / or TNF-f3 (Thl cells); ratio of -4, -5 ratio than -13 (Th2 cells), of 9 (of 9 ) than -17 (Thl7), of -22 (Th22), or other inflammatory cytokines. 其他效应性T细胞的细胞因子分泌谱是本领域技术人员已知的。 Other cytokine secreting effector T cells spectrum is known to the skilled person. 基于对各种效应性T 细胞群体的鉴定有用的表面标记的了解,本领域技术人员能够鉴定并枚举一个异质细胞群体中(例如在处于培养物中的一群细胞中或在获得自受试者的一群细胞中)的效应性T细胞。 Useful surface markers based on understanding of the identification of various effector T-cell population, the skilled in the art to identify and enumerate a heterogeneous cell population (e.g., in the culture of a population of cells obtained from the subject, or by a group of cells) effector T cells. 在其他实施例中,本领域技术人员可W查找细胞调亡、特异性T细胞(如CD4+或CD8巧细胞)的损失、或CD69活化,W便评定在此提供的运些组合物的功能。 In other embodiments, those skilled in the art may be W look apoptosis, loss of specific T cells (e.g. CD4 + or CD8 clever cells) or CD69 activation, W will assess the functions provided by the operation of these compositions.

[0143] 根据本发明,可W使用各种各样的合成纳米载体。 [0143] According to the present invention, it can be synthesized using a variety of W nanocarriers. 在一些实施例中,合成纳米载体是球体或球状体。 In some embodiments, synthetic nanocarriers are spheres or spheroids. 在一些实施例中,合成纳米载体是扁平的或盘状的。 In some embodiments, synthetic nanocarriers are flat or disk. 在一些实施例中,合成纳米载体是立方体或立方形的。 In some embodiments, synthetic nanocarriers cubic or cuboidal. 在一些实施例中,合成纳米载体是卵形或楠圆形的。 In some embodiments, synthetic nanocarriers Nan is oval or circular. 在一些实施例中,合成纳米载体是圆柱体、锥体、或锥形。 In some embodiments, synthetic nanocarriers is a cylinder, a cone, or a cone.

[0144] 在一些实施例中,希望使用在大小、形状、和/或构成方面较一致的一个群体的合成纳米载体,使得每一合成纳米载体具有相似特性。 [0144] In some embodiments, it is desirable to use in the size, shape, and / or the composition of more consistent synthetic nanocarriers a group, such that each synthetic nanocarriers having similar characteristics. 例如,基于合成纳米载体的总数,至少80%、至少90%、或至少95%的运些合成纳米载体可W具有归属在运些合成纳米载体的平均直径或平均尺寸的5%、10 %或20%之内的最小尺寸或最大尺寸。 For example, the total number of synthetic nanocarriers based, at least 80%, at least 90%, or at least 95% of the transport of these synthetic nanocarriers can be W with 5% attributable to the operation of these average diameter or average size of the synthetic nanocarriers, 10%, or minimum size or the maximum size within 20%. 在一些实施例中,一个群体的合成纳米载体可W在大小、形状、和/或构成方面是不均匀的。 In some embodiments, synthetic nanocarriers W may be a group of size, shape, and / or the composition of non-uniform.

[0145] 合成纳米载体可W是实屯、的或空屯、的,并且可W包含一个或多个层。 [0145] W is a synthetic nanocarriers can be real Tun, village or empty, and W may contain one or more layers. 在一些实施例中,每一层相对于其他一层或多层具有独特的构成和独特的特性。 In some embodiments, each layer has a unique phase characteristics and unique configuration other layer or layers. 为了作为一个实例给出,合成纳米载体可W具有一个核/壳结构,其中该核是一个层(例如一个聚合物核)并且该壳是一个第二层(例如一个脂质双层或单层)。 As an example is given to, synthetic nanocarriers W may have a core / shell structure, wherein the core is a layer (e.g., a core polymer) and the second shell is a layer (e.g., a lipid bilayer or monolayer ). 合成纳米载体可W包括多个不同的层。 Synthetic nanocarriers W may comprise a plurality of different layers.

[0146] 在一些实施例中,合成纳米载体可W任选地包含一种或多种脂质。 [0146] In some embodiments, synthetic nanocarriers W may optionally comprise one or more lipids. 在一些实施例中,合成纳米载体可W包括一种脂质体。 In some embodiments, synthetic nanocarriers W may include one liposomes. 在一些实施例中,合成纳米载体可W包含一个脂质双层。 In some embodiments, synthetic nanocarriers W may comprise a lipid bilayer. 在一些实施例中,合成纳米载体可W包含一个脂质单层。 In some embodiments, synthetic nanocarriers W may comprise a lipid monolayer. 在一些实施例中,合成纳米载体可W包括一种微胶粒。 In some embodiments, synthetic nanocarriers W may include one micelles. 在一些实施例中,合成纳米载体可W包含一个核,该核包含被一个脂质层(例如,脂质双层、脂质单层等等)围绕的一种聚合物基质。 In some embodiments, synthetic nanocarriers W may comprise a core which comprises a polymer matrix is ​​surrounded by a lipid layer (e.g., a lipid bilayer, a lipid monolayer, etc.). 在一些实施例中,合成纳米载体可W包含被一个脂质层(例如,脂质双层、脂质单层等等)围绕的一个非聚合物核(例如,金属颗粒、量子点、陶瓷颗粒、骨颗粒、病毒颗粒、蛋白质、核酸、碳水化合物等等)。 In some embodiments, synthetic nanocarriers may be W comprises a lipid layer (e.g., a lipid bilayer, lipid monolayer, etc.) a non-polymer core surrounded by (e.g., metal particles, quantum dots, the ceramic particles , bone particles, viral particles, proteins, nucleic acids, carbohydrates, etc.).

[0147] 在其他实施例中,合成纳米载体可W包含金属颗粒、量子点、陶瓷颗粒等等。 [0147] In other embodiments, synthetic nanocarriers W may comprise metal particles, quantum dots, ceramic particles and the like. 在一些实施例中,非聚合物合成纳米载体是非聚合组分的一个聚集体,如金属原子(例如金原子)的一个聚集体。 In some embodiments, the non-polymeric carrier is a synthetic nano aggregates a polymerizable component, such as a metal atom aggregates (e.g. gold atoms).

[014引在一些实施例中,合成纳米载体可W任选地包含一种或多种两亲实体。 [014] In some embodiments, primers, synthetic nanocarriers W may optionally comprise one or more amphiphilic entities. 在一些实施例中,两亲实体可W促进具有增加的稳定性、改进的均匀性、或增加的粘度的合成纳米载体的产生。 In some embodiments, an amphiphilic entity may facilitate W has increased stability, resulting synthetic nanocarriers improved uniformity, or increased viscosity. 在一些实施例中,两亲实体可W与一个脂质膜(例如,脂质双层、脂质单层等等) 的内表面结合。 In some embodiments, W may be bonded amphiphilic entity inner surface of a lipid membrane (e.g., a lipid bilayer, lipid monolayer, etc.). 在本领域中已知的许多两亲实体可W适用于制造根据本发明的合成纳米载体。 Many amphiphilic entity known in the art may be suitable for manufacturing the W synthetic nanocarriers of the present invention. 此类两亲实体包括但不限于:憐酸甘油醋类;憐脂酷胆碱类;二栋桐酷憐脂酷胆碱(DPPC);二油締基憐脂酷基乙醇胺(DOPE);二油締基氧丙基Ξ乙基锭(D0TMA);二油酷基憐脂酷胆碱;胆固醇;胆固醇醋;二酷基甘油;二酷基甘油班巧酸醋;二憐脂酷基甘油(DPPG); 十六烧醇;脂肪醇类,如聚乙二醇(PEG);聚氧乙締-9-月桂基酸;一种表面活性脂肪酸,如栋桐酸或油酸;脂肪酸类;脂肪酸甘油单醋类;脂肪酸甘油二醋类;脂肪酸酷胺类;脱水山梨糖醇Ξ油酸醋(沒pan@85)甘氨胆酸醋;脱水山梨糖醇单月桂酸醋(没pan@20);聚山梨醇醋20(Tween敏20);聚山梨醇醋60(Twee田嚷60);聚山梨醇醋65(Tween®65);聚山梨醇醋80(lWeen®80);聚山梨醇醋85(Tween够85);聚氧乙締单硬脂酸醋;表面活性素;泊洛沙姆;一种脱水山梨糖醇脂肪酸醋,如脱水山梨糖醇Ξ油酸醋;卵憐脂;溶血卵憐脂;憐脂酷丝氨酸;憐脂酷肌醇;銷憐脂;憐脂酷 Such amphiphilic entities include, but are not limited to: pity nitroglycerin vinegar; pity aliphatic cool choline; Tong two aliphatic cool cool pity choline (of DPPC); dioleyl associative aliphatic group pity cool ethanolamine (DOPE); two oil-based associative oxopropyl Ξ ethyl ingot (D0TMA); pity lipid dioleyl cool cool choline group; cholesterol; cholesterol vinegar; two cool glycerol; glycerol cool two classes Qiao vinegar; two aliphatic cool pity glycerol ( DPPG); sixteen burning alcohol; fatty alcohols such as polyethylene glycol (PEG); polyoxyethylene-9-lauryl associative acid; a surface active fatty acid, such as oleic acid or eleostearic buildings; fatty acids; fatty acid mono- vinegar; vinegar diglycerides of fatty acids; fatty amines cool; sorbitan alcohols Ξ oil vinegar (did not pan @ 85) glycocholate vinegar; sorbitan monolaurate alcohol vinegar (did not pan @ 20) ; vinegar polysorbate 20 (Tween 20 Min); vinegar polysorbate 60 (Twee cried field 60); vinegar polysorbate 65 (Tween®65); vinegar polysorbate 80 (lWeen®80); polysorbate vinegar 85 (Tween 85 enough); polyoxyethylene mono association acetate stearate; surfactin; poloxamer; a dehydrating sorbitan fatty acid vinegar, sugar alcohols such as sorbitan Ξ oil vinegar; egg lipid pity; hemolysis pity egg fat; grease pity cool serine; cool inositol lipid pity; pity pin aliphatic; aliphatic cool pity 醇胺(脑憐脂);屯、憐脂;憐脂酸;脑巧脂;双十六烧基憐酸醋;二栋桐酷憐脂酷甘油;硬脂酷胺;十二烧胺;十六烧胺;乙酷基栋桐酸醋;藍麻油酸甘油醋;十八酸十六烷基醋;肉豆違酸异丙醋;四下酪醒(tyloxapol);聚(乙二醇)5000- 憐脂酷乙醇胺;聚(乙二醇)400-单硬脂酸醋;憐脂;具有高表面活性剂特性的合成的和/或天然的洗涂剂;脱氧胆酸醋;环糊精;离液序列高的盐;离子对试剂;W及它们的组合。 Alcohol amines (aliphatic brain pity); Tun, aliphatic pity; pity fatty acid; Qiao brain lipid; burned dihexadecyl group pity vinegar; Tong two aliphatic cool cool pity glycerol; cool stearyl amine; twelve burn amine; ten six burning amine; yl acetate cool buildings Kazuto vinegar; glycerol, acetic acid, sesame oil blue; stearic acid cetyl acetate; myristoyl illegally acid isopropyl acetate; awake looked casein (Tyloxapol); poly (ethylene glycol) 5000 - cool pity aliphatic amine; poly (ethylene glycol) 400- vinegar monostearate; pity fats; synthetic and / or natural wash varnish having a high surface active agent properties; deoxycholate vinegar; cyclodextrin; chaotropic salt sequence; ion pair reagent; W is and combinations thereof. 两亲实体组分可W是不同两亲实体的混合物。 W amphiphilic entity component may be mixtures of different amphiphilic entities. 本领域普通技术人员将认识到,运是具有表面活性剂活性的物质的示例性的、而不是全面的清单。 Those of ordinary skill in the art will recognize that the operation is an exemplary active substance is a surfactant, not a comprehensive list. 任何两亲实体可W用于有待根据本发明使用的合成纳米载体的生产中。 Any amphiphilic entity may be used for the production of synthesis W nanocarriers in accordance with the present invention.

[0149] 在一些实施例中,合成纳米载体可W任选地包含一种或多种碳水化合物。 [0149] In some embodiments, synthetic nanocarriers W may optionally comprise one or more carbohydrates. 碳水化合物可W是天然的或合成的。 W carbohydrate is natural or synthetic. 碳水化合物可W是衍生的天然碳水化合物。 W is a derivatized carbohydrates natural carbohydrate. 在某些实施例中, 碳水化合物包含单糖或二糖,包括但不限于:葡萄糖、果糖、半乳糖、核糖、乳糖、薦糖、麦芽糖、海藻糖、纤维二糖(cel Aiose)、甘露糖、木糖、阿拉伯糖、葡糖醒酸、半乳糖醒酸、甘露糖醒酸、葡糖胺、半乳糖胺、W及神经氨酸。 In certain embodiments, the carbohydrate comprises a monosaccharide or disaccharide, including but not limited to: glucose, fructose, galactose, ribose, lactose, recommended, maltose, trehalose, cellobiose (cel Aiose), mannose , xylose, arabinose, wake gluconic acid, galacturonic acid wake, wake acid mannose, glucosamine, galactosamine, W, and neuraminic acid. 在某些实施例中,碳水化合物是一种多糖,包括但不限于:支链淀粉、纤维素、微晶纤维素、径丙基甲基纤维素化PMC)、径基纤维素化C)、甲基纤维素(MC)、葡聚糖、环葡聚糖、糖原、径乙基淀粉、卡拉胶、多聚糖(glycon)、直链淀粉、壳聚糖、N,0-簇甲基壳聚糖、藻胶和海藻酸、淀粉、甲壳质、菊糖、魔芋、葡萄甘露聚糖(glucommannan)、石耳素、肝素、透明质酸、凝胶多糖、W及黄原胶。 In certain embodiments, the carbohydrate is a polysaccharide, including but not limited to: pullulan, cellulose, microcrystalline cellulose, methylcellulose diameter of the PMC), the diameter of the cellulose group C), methylcellulose (MC), dextran, cyclodextran, glycogen, diameter ethyl starch, carrageenan, polysaccharides (Glycon), amylose, chitosan, N, 0- cluster methyl chitosan, algin and alginic acid, starch, chitin, inulin, konjac, glucomannan (glucommannan), pustulan, heparin, hyaluronic acid, curdlan, W, and xanthan gum. 在实施例中,运些发明的合成纳米载体并不包含(或特别排除)碳水化合物,如多糖。 In an embodiment, the invention these synthetic nanocarriers shipped does not include (or specifically excluded) carbohydrates, such as polysaccharides. 在某些实施例中,该碳水化合物可W包含一种碳水化合物衍生物,如一种糖醇,包括但不限于:甘露醇、山梨醇、木糖醇、赤薛糖醇、麦芽糖醇、W及乳糖醇。 In certain embodiments, the carbohydrate can include one W carbohydrate derivative, such as a sugar alcohol, including but not limited to: mannitol, sorbitol, xylitol, erythritol Xue sorbitol, maltitol, and W lactitol.

[0150] 在一些实施例中,合成纳米载体可W包含一种或多种聚合物。 [0150] In some embodiments, synthetic nanocarriers W may comprise one or more polymers. 在一些实施例中,运些合成纳米载体包含一种或多种聚合物,该聚合物是一种非甲氧基封端的普朗尼克聚合物。 In some embodiments, some synthetic nanocarriers transport comprises one or more polymers, the polymer is a non-methoxy-terminated Pluronic polymers. 在一些实施例中,构成运些合成纳米载体的至少1%、2%、3%、4%、5%、10%、15%、 20%、25%、30%、35%、40%、45%、50%、55%、60%、65%、70%、75%、80%、85%、90%、 95 %、97 %、或99 % (重量/重量)的运些聚合物是非甲氧基封端的普朗尼克聚合物。 In some embodiments, constituting the transport of these synthetic nanocarriers at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, or 99% (w / w) of non-operation of these polymers methoxy-terminated Pluronic polymers. 在一些实施例中,构成运些合成纳米载体的所有运些聚合物是非甲氧基封端的普朗尼克聚合物。 In some embodiments, some synthetic nanocarriers constituting transport all of these polymers are transported methoxy-terminated Pluronic polymers. 在一些实施例中,运些合成纳米载体可W包含一种或多种聚合物,该聚合物是非甲氧基封端的聚合物。 In some embodiments, some synthetic nanocarriers transport W may comprise one or more polymers, non-methoxy-terminated polymer of the polymer. 在一些实施例中,构成运些合成纳米载体的至少1%、2%、3%、4%、5%、10%、 15%、20%、25%、30%、35%、40%、45%、50%、55%、60%、65%、70%、75%、80%、85%、 90%、95%、97%、或99% (重量/重量)的运些聚合物是非甲氧基封端的聚合物。 In some embodiments, constituting the transport of these synthetic nanocarriers at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, or 99% (w / w) of non-operation of these polymers methoxy-terminated polymer. 在一些实施例中,构成运些合成纳米载体的所有运些聚合物是非甲氧基封端的聚合物。 In some embodiments, the polymer constituting the synthetic nanocarriers transport some of all of these polymers are transported methoxy terminated. 在一些实施例中,运些合成纳米载体包含一种或多种聚合物,运些聚合物不包含普朗尼克聚合物。 In some embodiments, some synthetic nanocarriers transport comprises one or more polymers, these polymers do not contain transport pluronic polymer. 在一些实施例中,构成运些合成纳米载体的至少1%、2%、3%、4%、5%、10%、15%、20%、25%、 30%'35%'40%'45%'50%'55%'60%'65%'70%'75%'80%'85%'90%'95%'97%、或99% (重量/重量)的运些聚合物不包含普朗尼克聚合物。 In some embodiments, some synthetic nanocarriers transport constituting at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30% '35% '40% ' '50 '55% 45%%% '70 '65 '60%%% '85 '80 '75%%% '97 '95 '90%%, or 99% (wt / wt) of these polymers does not run Pluronic polymers comprising. 在一些实施例中,构成运些合成纳米载体的所有运些聚合物不包含普朗尼克聚合物。 In some embodiments, some synthetic nanocarriers transport constituting all these polymers do not contain transport pluronic polymer. 在一些实施例中,可W由包被层(例如脂质体、脂质单层、微胶粒等)环绕运样一种聚合物。 In some embodiments, W can be transported by a sample coated with a polymer layer (for example, liposomes, lipid monolayers, micelles, etc.) surrounds. 在一些实施例中,合成纳米载体的不同成分可W与该聚合物偶联。 In some embodiments, different compositions of synthetic nanocarriers W may be conjugated to the polymer.

[0151] 可W通过多种方法中的任一种使运些免疫抑制剂和/或抗原与运些合成纳米载体偶联。 [0151] W may be by any of a variety of methods to make transport more immunosuppressant and / or transport of these antigens conjugated synthetic nanocarriers. 通常,该偶联可W是运些免疫抑制剂和/或抗原与运些合成纳米载体之间键合的结果。 Typically, the coupling can be more immunosuppressant W is transported and / or antigen to transport between the results of these synthetic nanocarriers bonded. 此键合可W导致运些免疫抑制剂和/或抗原附接至运些合成纳米载体的表面上和/或包含(封装)在运些合成纳米载体内。 This bonding operation may cause some immunosuppressive W and / or the antigen is attached to the upper surface of the operation of some synthetic nanocarriers, and / or comprising (package) in the transport of these synthetic nanocarrier. 然而,在一些实施例中,运些免疫抑制剂和/或抗原由于运些合成纳米载体的结构而被合成纳米载体封装,而不是键合至合成纳米载体上。 However, in some embodiments, some immunosuppressive shipped and / or transported antigen since synthetic nanocarriers such structures are synthetic nanocarriers package, rather than bonded to the synthetic nanocarrier. 在多个优选的实施例中,合成纳米载体包含如在此所提供的一种聚合物,并且运些免疫抑制剂和/ 或抗原与聚合物偶联。 In preferred embodiments, the nanocarrier comprising a polymer synthesized as provided herein, and some immunosuppressive transport and / or the antigen conjugated to a polymer.

[0152] 当由于运些免疫抑制剂和/或抗原与合成纳米载体之间的键合而发生偶联时,该偶联可W经由一个偶联部分而发生。 [0152] When due to the operation between these immunosuppressive and / or antigen bonded synthetic nanocarriers coupling occurs, the coupling may be via a coupling portion W occurs. 一个偶联部分可W是通过其使免疫抑制剂和/或抗原键合至合成纳米载体上的任何部分。 W is a coupling moiety may be by which the immunosuppressive and / or antigens bound to any part of the synthetic nanocarriers. 此类部分包括共价键(如一个酷胺键或醋键)、W及使免疫抑制剂和/或抗原键合(共价地或非共价地)至该合成纳米载体上的单独的分子。 Such moieties include covalent bonds (such as a key or vinegar cool amine bond), W and immune inhibitor and / or antigen bound (covalently or non-covalently) to the synthetic nanocarriers on a separate molecule . 此类分子包括连接物或聚合物或其单元。 Such molecules include linker or polymer units thereof. 例如,该偶联部分可W包含免疫抑制剂和/或抗原静电结合至其上的一种带电荷的聚合物。 For example, the coupling portion may comprise an immunosuppressant W and / or electrostatically bound to the antigen on which one charged polymer. 作为另一个实例,该偶联部分可W包含它共价结合至其上的一种聚合物或其单元。 As another example, the coupling portion may comprise W it covalently bound to a polymer or a unit thereon.

[0153] 在多个优选的实施例中,运些合成纳米载体包含如在此所提供的一种聚合物。 [0153] In preferred embodiments, the transport of these synthetic nanocarrier comprising a polymer as provided herein. 运些合成纳米载体可W是完全聚合的或它们可W是聚合物与其他材料的混合物。 These synthetic nanocarriers can be transported W are fully polymerized or they may be mixtures of polymers and W are other materials.

[0154] 在一些实施例中,具有合成纳米载体的运些聚合物结合W形成一种聚合物基质。 [0154] In some embodiments, these polymers have transport binding synthetic nanocarriers W form a polymer matrix. 在运些实施例中的一些中,一种组分(如一种免疫抑制剂或抗原)可W与该聚合物基质中的一种或多种聚合物共价结合。 In some embodiments some of the transport, one component (e.g., an immunosuppressant or antigen) W embodiment may be combined with one of the polymer matrix in a covalent or more polymers. 在一些实施例中,共价结合是由一个连接物介导的。 In some embodiments, the covalent binding is mediated by a linker. 在一些实施例中,一种组分可W与该聚合物基质中的一种或多种聚合物非共价结合。 In some embodiments, one component of W may be combined with the polymer matrix of one or more polymers non-covalently. 例如,在一些实施例中,一种组分可W被封装在一种聚合物基质之内、被聚合物基质围绕、和/或分散在整个聚合物基质中。 For example, in some embodiments, W can be one component encapsulated within a polymer matrix, the polymer matrix is ​​surrounded by, and / or dispersed throughout the polymer matrix. 可替代地或另外地,一种组分可W通过疏水相互作用、电荷相互作用、范德华力等等与一种聚合物基质中的一种或多种聚合物结合。 Alternatively or additionally, one component may be W through hydrophobic interactions, charge interactions, Van der Waals forces and the like in combination with a polymer matrix of one or more polymers. 用于从其形成聚合物基质的各种各样的聚合物和方法是常规已知的。 Polymers and various methods for forming therefrom the polymer matrix are conventionally known.

[0155] 聚合物可W是天然的或非天然的(合成的)聚合物。 [0155] W is a polymer of natural or non-natural (synthetic) polymer. 聚合物可W是均聚物或包含两种或更多种单体的共聚物。 W is a polymer can be a homopolymer or a copolymer comprising two or more monomers. 在序列方面,共聚物可W是随机的、嵌段的,或包含随机序列与嵌段序列的组合。 In sequence, W may be random copolymers, block, or a combination of random sequence with the block sequence. 典型地,根据本发明的聚合物是有机聚合物。 Typically, the polymer according to the present invention is an organic polymer.

[0156] 在一些实施例中,聚合物包含一种聚醋、聚碳酸醋、聚酷胺、或聚酸、或其单元。 [0156] In some embodiments, the polymer comprises one polyester, polycarbonate, polyethylene amine cool, or dimer acid, or a unit. 在其他实施例中,聚合物包含聚(乙二醇KPEG)、聚丙二醇、聚(乳酸)、聚(乙醇酸)、聚(乳酸- 乙醇酸)共聚物、或一种聚己酸内醋、或其单元。 In other embodiments, the polymer comprises a poly (ethylene glycol KPEG), polypropylene glycol, poly (lactic acid), poly (glycolic acid), poly (lactic acid - glycolic acid) copolymer, a poly-caprolactone acid or vinegar, or a unit. 在一些实施例中,优选的是该聚合物是可生物降解的。 In some embodiments, it is preferred that the polymer is biodegradable. 因此,在运些实施例中,优选的是如果该聚合物包含一种聚酸(如聚(乙二醇)或聚丙二醇或其单元),则该聚合物包含一种聚酸和一种可生物降解的聚合物的嵌段共聚物, 使得该聚合物是可生物降解的。 Thus, in some embodiments, transportation, it is preferred if the polymer comprises a poly acids (e.g., poly (ethylene glycol) or poly propylene glycol or means), the polymer comprises an acid and a poly A biodegradable a block copolymer, such that the polymer is biodegradable. 在其他实施例中,该聚合物不仅仅包含一种聚酸或其单元, 如聚(乙二醇)或聚丙二醇或其单元。 In other embodiments, the polymer comprises a poly acid or just units, such as poly (ethylene glycol) or polyethylene glycol, or a unit.

[0157] 适合用于本发明的聚合物的其他实例包括但不限于:聚乙締、聚碳酸醋(例如聚(1,3 -二囉烧-2酬))、聚酢(例如聚(癸二酸酢))、聚丙基延胡索酸醋(polypropylfumerate)、聚酷胺(例如聚己内酷胺)、聚缩醒、聚酸、聚醋(例如聚丙交醋、聚乙交醋、丙交醋-乙交醋共聚物、聚己酸内醋、多径基酸(例如聚(0-径基烧酸醋)、聚(原酸醋)、聚氯基丙締酸醋、聚乙締醇、聚氨醋、聚憐腊、聚丙締酸醋、聚甲基丙締酸醋、聚脈、聚苯乙締、W及聚胺、聚赖氨酸、聚赖氨酸-PEG共聚物、W及聚化締亚胺)、聚化締亚胺)-PEG共聚物。 [0157] Other examples of suitable polymers for the present invention include, but are not limited to: polyethylene association, polycarbonates (e.g., poly (1,3 - La burn -2 paid)), poly Health (e.g., poly (decyl Health diacid)), polypropylene fumarate group vinegar (polypropylfumerate), poly cool amine (e.g. polycaprolactone cool amine), polycondensation awake, polylactic acid, polyesters (e.g., polylactide vinegar, vinegar polyglycolides, lactide vinegar - vinegar lactide copolymers, polycaprolactone vinegar, multipath acids (e.g., poly (o-burn-yl diameter vinegar), poly (ortho vinegar), polychlorotrifluoroethylene propionic associated vinegar, associative polyethylene glycol, polyethylene ammonia, vinegar, pity polyethylene wax, polypropylene associative vinegar, vinegar association polymethyl methacrylate, polyurea, polystyrene association, W, and polyamines, polylysine, polylysine -PEG copolymers, W, and poly of the association), poly imines of association) -PEG copolymers.

[015引在一些实施例中,根据本发明的聚合物包括在21C.FR§177.2600下已经由美国食品与药品管理局(FDA)批准用于人的聚合物,包括但并不局限于聚醋(例如聚乳酸、聚(乳酸乙醇酸共聚物)、聚己内醋、聚戊内醋、聚(1,3-二囑酬));聚酢(例如聚(癸二酸酢));聚酸(例如聚乙二醇);聚氨醋;聚甲基丙締酸醋;聚丙締酸醋;W及聚氯基丙締酸醋。 [015] In some embodiments cited, according to the present invention comprises a polymer 21C.FR§177.2600 has the U.S. Food and Drug Administration (FDA) approved for human use by the polymers, including, but not limited to, polyester (e.g. polylactic acid, poly (lactic-glycolic acid), polycaprolactone vinegar, vinegar amyl cohesion, poly (1,3-Well paid)); poly Health (e.g., poly (sebacic acid Health)); poly acid (e.g., polyethylene glycol); polyurethane vinegar; polymethyl methacrylate associative vinegar; polypropylene association vinegar; W is associated vinegar and propionic polychloroprene.

[0159] 在一些实施例中,聚合物可W是亲水性的。 [0159] In some embodiments, the polymer may be W is hydrophilic. 例如,聚合物可W包含阴离子基团(例如憐酸根、硫酸根、簇酸根);阳离子基团(例如,季胺基团);或极性基团(例如,径基基团、硫醇基团、胺基团)。 For example, the polymer can contain anionic group W (e.g. pity, sulphate, and phosphate cluster); cationic groups (e.g., quaternary amine groups); or a polar group (e.g., diameter group, a thiol group group, amine group). 在一些实施例中,包含一种亲水性聚合物基质的合成纳米载体在该合成纳米载体内产生亲水环境。 In some embodiments, the polymer matrix comprises one hydrophilic synthetic nanocarriers hydrophilic environment produced in the synthetic nanocarrier. 在一些实施例中,聚合物可W是疏水性的。 In some embodiments, the polymer may be W is hydrophobic. 在一些实施例中,包含疏水性聚合物基质的合成纳米载体在该合成纳米载体内产生疏水环境。 In some embodiments, the nanocarrier comprising a hydrophobic synthetic polymer matrix is ​​generated within the hydrophobic environment synthetic nanocarrier. 聚合物亲水性或疏水性的选择可W影响被结合(例如偶联)在合成纳米载体之内的材料的性质。 Hydrophilic or hydrophobic polymers of choice may be W affect binding properties (e.g. conjugation) within the composite material of the nanocarrier.

[0160] 在一些实施例中,聚合物可W用一个或多个部分和/或官能团修饰。 [0160] In some embodiments, the polymer may be one or more portions with W and / or functional group modifications. 根据本发明, 可W使用各种各样的部分或官能团。 According to the present invention, W using various moieties or functional groups. 在一些实施例中,可W用聚乙二醇(PEG)、用碳水化合物、和/或用衍生自多糖类的非环状聚缩醒来修饰聚合物(Papisov(己比索夫),2001,ACS Symposium Series,786:301)。 In some embodiments, W can be a polyethylene glycol (PEG), with a carbohydrate, and / or with polysaccharides derived from non-cyclic-modified polymers of polycondensation wake (Papisov (Bi Suofu hexyl), 2001 , ACS Symposium Series, 786: 301). 可W使用Gref (格里夫)等人的美国专利号5543158、或Von Amlrian(冯安德里安)等人的WO公开W02009/051837中的全部传授内容进行某些实施例。 W may be used Gref (Grieve) et al., U.S. Patent No. 5,543,158, or Von Amlrian (von Andrian) et al., The entire disclosure of WO W02009 / 051837 teachings for certain embodiments.

[0161] 在一些实施例中,可W用脂质或脂肪酸基团修饰聚合物。 [0161] In some embodiments, W may be a group with a lipid or fatty acid modified polymers. 在一些实施例中,脂肪酸基团可W是下酸、己酸、辛酸、癸酸、月桂酸、肉豆違酸、栋桐酸、硬脂酸、花生酸、山斋酸、或廿四烧酸中的一种或多种。 In some embodiments, W is a fatty acid group may be an acid, caproic acid, caprylic acid, capric acid, lauric acid myristoyl violation, buildings eleostearic acid, stearic acid, arachidic acid, behenic acid, vegetarian, or round-burning one or more acids. 在一些实施例中,脂肪酸基团可W是栋桐締酸、油酸、异油酸、亚麻酸、α-亚麻酸、丫-亚麻酸、花生四締酸、二十碳締酸、花生四締酸、二十碳五締酸、二十二碳六締酸、或芥酸中的一种或多种。 In some embodiments, W is a fatty acid group may be associated ridge eleostearic acid, oleic acid, vaccenic acid, linoleic acid, linolenic acid alpha], Ah - linolenic acid, arachidonic acid association, association eicosenoic acid, arachidonic Parties acid, eicosapentaenoic acid association, association docosahexaenoic acid, or one or more of erucic acid.

[0162] 在一些实施例中,聚合物可W是聚醋,包括共聚物,运些共聚物包括乳酸和乙醇酸单元(例如聚(乳酸乙醇酸共聚物)和聚(丙交醋乙交醋共聚物)),在此统称为"PLGA";W及包括乙醇酸单元的均聚物,在此称为"PGA",W及乳酸单元(例如聚乳酸、聚-D-乳酸、聚- D,k乳酸、聚-k丙交醋、聚-D-丙交醋、W及聚-D,k丙交醋),在此统称为"PLA"。 [0162] In some embodiments, W is a polyester polymer, comprising copolymers, these copolymers comprising a transport unit lactic and glycolic acids (e.g., poly (lactic-co-glycolic acid) and poly (lactide glycolide vinegar vinegar copolymer)), referred to herein as "PLGA"; W, and comprises a homopolymer of glycolic acid units, referred to herein as "PGA", W and lactic acid unit (e.g. polylactic acid, -D- acid, poly - D , k lactic acid, poly-lactide -k vinegar, vinegar poly-lactide -D-, W, and poly--D, k vinegar-lactide), referred to herein as "PLA". 在一些实施例中,示例性的聚醋包括,例如多径基酸;PEG共聚物和丙交醋与乙交醋的共聚物(例如PLA-PEG共聚物、PGA-阳G共聚物、PLGA-PEG共聚物),W及它们的衍生物)。 In some embodiments, the exemplary polyester include, for example, multipath acids; PEG copolymer and a copolymer of lactide and glycolide vinegar vinegar (e.g., PLA-PEG copolymers, copolymers of PGA-Yang G, the PLGA- PEG copolymer), W and derivatives thereof). 在一些实施例中, 聚醋包括例如聚(己内醋)、聚(己内醋)-PEG共聚物、聚化-丙交醋赖氨酸)共聚物、聚(丝氨酸醋)、聚(4-径基A-脯氨酸醋)、聚[α-(4-氨基下基)A-乙醇酸]、W及其衍生物。 In some embodiments, comprises a polyester such as poly (caprolactone vinegar), poly (caprolactone vinegar) -PEG copolymers, of poly - lysine vinegar lactide), poly (serine acetate), poly (4 - proline A- diameter yl vinegar), poly [alpha] (at 4-yl) A- glycolic acid], W is and derivatives thereof.

[0163] 在一些实施例中,聚合物可W是化GAdPLGA是一种生物相容的并且生物可降解的乳酸和乙醇酸的共聚物,并且多种形式的PLGA特征在于乳酸:乙醇酸的比率。 [0163] In some embodiments, W is a polymer of GAdPLGA is a biocompatible and biodegradable copolymer of lactic acid and glycolic acid, and various forms of PLGA comprising lactic: glycolic acid ratio . 乳酸可^是心乳酸、D-乳酸、或D,k乳酸。 Lactic acid can heart ^, D- lactic acid, or D, k lactic acid. 可W通过改变乳酸:乙醇酸的比率调整PLGA的降解速率。 W may be changed by lactic acid: glycolic acid ratio adjusted rate of degradation of PLGA. 在一些实施例中,根据本发明的将使用的PLGA特征在于大约85:15、大约75:25、大约60:40、大约50 :50、大约40:60、大约25:75、或者大约15:85的乳酸:乙醇酸比率。 In some embodiments, in accordance with the PLGA used in the present invention is characterized in that about 85:15, about 75:25, about 60:40, about 50: 50, about 40:60, about 25:75, or about 15: 85 lactic acid: glycolic acid ratio.

[0164] 在一些实施例中,聚合物可W是一种或多种丙締酸聚合物。 [0164] In some embodiments, W is a polymer of one or more propionic acid associative polymer. 在某些实施例中,丙締酸聚合物包括,例如丙締酸和甲基丙締酸的共聚物、甲基丙締酸甲醋共聚物、甲基丙締酸乙氧基乙醋、甲基丙締酸氯基乙基醋、甲基丙締酸氨基烷基醋共聚物、聚(丙締酸)、聚(甲基丙締酸)、甲基丙締酸烷基酷胺共聚物、聚(甲基丙締酸甲醋)、聚(甲基丙締酸酸酢)、甲基丙締酸甲醋、聚甲基丙締酸醋、聚(甲基丙締酸甲醋)共聚物、聚丙締酷胺、甲基丙締酸氨基烷基醋共聚物、甲基丙締酸缩水甘油醋共聚物、聚腊基丙締酸醋、W及包括一种或多种W上聚合物的组合。 In certain embodiments, the associative propionic acid polymers include, for example, a copolymer of propionic acid and methacrylic associated associative acid, methacrylic acid methyl ester copolymers association, the association methylpropanesulfonic acid ethoxyethyl acetate, methyl propionic acid chloride associative ethyl acetate, methyl propionic acid aminoalkyl association acetate copolymer, poly (propyl association acid), poly (methyl propionic acid association), methacrylic acid alkyl association cool amine copolymer, poly (meth association propionic acid methyl ester), poly (methyl-prop-sour association of Health), the association methylpropanesulfonic acid methyl ester, polymethyl methacrylate associative vinegar, poly (meth association propionic acid methyl ester) copolymer , polypropoxylated amine associated cool, methacrylic acid aminoalkyl association acetate copolymers, methacrylic acid, glycidyl acetate associative copolymer, polyethylene wax propionic associated vinegar, the W and W comprises one or more polymers combination. 该丙締酸聚合物可W包含具有低含量的季锭基团的丙締酸醋和甲基丙締酸醋的完全聚合的共聚物。 Propionic acid The associative polymer may comprise ingot W quaternary group with a low content of fully polymerized copolymers of propionic and methacrylic associated vinegar association of vinegar.

[0165] 在一些实施例中,聚合物可W是阳离子型聚合物。 [0165] In some embodiments, the polymer may be W is a cationic polymer. 通常,阳离子型聚合物能够缩合和/或保护核酸(例如DNA、或它的衍生物)的带负电的链。 Typically, the cationic polymer can be a condensation and / or protect the nucleic acid (e.g. the DNA, or a derivative thereof) is negatively charged strand. 含有胺的聚合物(例如聚(赖氨酸) (Zauner(泽纳)等人,1998,先进药物递送综述,30:97; W及Kabanov(卡己诺夫)等人,1995, 生物共辆化学,6:7)、聚(亚乙基亚胺)(阳I;Boussif (波希夫)等人,1995,美国科学院院刊, 1995,92:7297)、W及聚(酷胺胺)树枝状聚合物化址owska(库科斯卡)-La化llo(拉塔罗)等人,1996,美国科学院院刊,93 :4897 ;Tang(唐)等人,1996,生物共辆化学,7:703 及化ensler(亨斯勒)等人,1993,生物共辆化学,4:372)在生理抑下是带正电的,在多种细胞系中与核酸形成离子对,并且介导转染。 Human polymer containing an amine (e.g., poly (lysine) (Zauner (zena) et al., 1998, advanced drug delivery Review, 30: 97; W and Kabanov (Nove card hexyl) et al., 1995, bioconjugates vehicle Chemistry, 6: 7), poly (ethylene imine) (Yang I; Boussif (Bo Xifu) et al., 1995, Proc Natl Acad Sci USA, 1995,92: 7297), W, and poly (cool amine) dendrimers of the site owska (Kukesika) -La of llo (Lata Luo) et al., 1996, Proc Natl Acad Sci USA, 93: 4897; Tang (Tang) et al., 1996, total units of biological chemistry, 7: and 703 of Ensler (Hensler) et al., 1993, bioconjugate chemistry vehicles, 4: 372) at physiological suppression is positively charged, forming ion pairs with nucleic acids in various cell lines, and transfection mediated . 在多个实施例中,运些发明的合成纳米载体可W不包含(或可W排除)阳离子型聚合物。 In various embodiments, some synthetic nanocarriers transported invention may not comprise W (or W excluded) cationic polymer.

[0166] 在一些实施例中,聚合物可W是带有阳离子侧链的可降解聚醋(Putnam(普特南) 等人,1999,大分子,32:3658 ;Barrera(己雷拉)等人,1993,美国化学会志,115:11010 ;Kwon (权)等人,1989,大分子,22:3250 ;Lim(林)等人,1999,美国化学会志,121:5633 及Zhou (周)等人,1990,大分子,23:3399)。 [0166] In some embodiments, W is a biodegradable polymer with a cationic side chain polyester (Putnam (Putnam) et al., 1999, macromolecular, 32: 3658; Barrera (Cabrera hexyl), etc. al., 1993, American chemical Society, 115: 11010; Kwon (right) et al., 1989, macromolecules, 22: 3250; Lim (Lin) et al., 1999, American chemical Society, 121: 5633 and Zhou (Zhou ) et al., 1990, macromolecules, 23: 3399). 运些聚合物的实例包括聚化-丙交醋k赖氨酸共聚物) (Barrera(己雷拉)等人,1993,美国化学会志,115:11010)、聚(丝氨酸醋)(Zhou(周)等人, 1990,大分子,23:3399)、聚(4-径基-心脯氨酸醋)(化tnam(普特南)等人,1999,大分子,32: 3658; W及Lim(林)等人,1999,美国化学会志,121:5633)、^及聚(4-径基-1^11氨酸醋) (Putnam(普特南)等人,1999,大分子,32 :3658 及Lim(林)等人,1999,美国化学会志, 121:5633)。 Examples of operation of these polymers include poly - lactide k acetate copolymer lysine) (Barrera (Herrera hexyl) et al., 1993, American Chemical Society, 115: 11010), poly (serine vinegar) (Zhou ( weeks) et al., 1990, macromolecular, 23: 3399), poly (4-yl-diameter - core proline vinegar) (of TNAM (Putnam) et al., 1999, macromolecular, 32: 3658; W and lim (Lin) et al., 1999, American chemical Society, 121: 5633), ^, and poly (4-1 ^ 11 diameter vinegar acid) (Putnam (Putnam) et al., 1999, macromolecules, 32: 3658 and Lim (Lin) et al., 1999, American chemical Society, 121: 5633).

[0167] 运些和其他聚合物的特性W及用于制备它们的方法在本领域中是熟知的(参见, 例如美国专利6,123,727; 5,804,178; 5,770,417; 5,736,372; 5,716,404;6,095,148;5, 837,752;5,902,599;5,696,175;5,514,378;5,512,600;5,399,665;5,019,379;5,010, 167;4,806,621;4,638,045; W及4,946,929;Wang(王)等人,2001,美国化学会志,123: 9480;Lim(林)等人,2001,美国化学会志,123:2460;Langer(朗格尔),2000,化学研究评述, 33:94; Langer (朗格尔),1999,控释杂志,62:7; W及Uhrich(乌利希)等人,1999,化学评论, 99:3181)。 [0167] These and other transport properties of the polymer, and W for their preparation are well known in the art (see, e.g. U.S. Patent No. 6,123,727; 5,804,178; 5,770,417; 5,736,372; 5,716,404; 6,095,148; 5, 837,752; 5,902,599; 5,696,175; 5,514,378; 5,512,600; 5,399,665; 5,019,379; 5,010, 167; 4,806,621; 4,638,045; W and 4,946,929; Wang (Wang) et al., 2001, American chemical Society, 123: 9480; Lim (Lin) et al., 2001 American chemical Society, 123: 2460; 2000, chemical Research Review Langer (Langer), 33:94; Langer (Langer), 1999, Journal of Controlled release, 62: 7; W and Uhrich (Uli Greek) et al., 1999, chemical reviews, 99: 3181). 更一般地,在Concise Encyclopedia of Polymer Science and Polymeric Amines and Ammonium Salts,Goethals(戈萨尔斯)编辑,培格曼出版社,1980中;在Principles of Polymerization by Odian,约翰•威利父子出版公司,第四版,2004中;在Allcock(阿尔库克)等人的Contemporary Polymer Qiemistry,Prentice-化11,1981 中;在Deming(德明)等人,1997,自然,390:386中;^及在美国专利6,506,577、6,632,922、6,686, 446、W及6,818,732中说明了用于合成某些适合的聚合物的多种方法。 More generally, of Polymer Science and Polymeric Amines and Ammonium Salts, Goethals (Zaragoza Wales) edited Concise Encyclopedia, Pergamon Press, in 1980; in Principles of Polymerization by Odian, • John Wiley & Sons, fourth edition, 2004; in Allcock (al Cook) et al., Contemporary Polymer Qiemistry, Prentice- of 11,1981; and in Deming (Deming) et al., 1997, Nature, 390: 386; ^ and in U.S. Patent No. 6,506,577,6,632,922,6,686, 446, W, and 6,818,732 illustrate various methods for the synthesis of some suitable polymers.

[0168] 在一些实施例中,聚合物可W是线型聚合物或分支聚合物。 [0168] In some embodiments, the polymer may be W is a linear polymer or branched polymer. 在一些实施例中,聚合物可W是树枝状化合物。 In some embodiments, W is a dendrimer polymer. 在一些实施例中,聚合物可W是基本上彼此交联的。 In some embodiments, W is a polymer can be crosslinked with each other substantially. 在一些实施例中,聚合物可W基本上不交联。 In some embodiments, the polymer may be W is not substantially crosslinked. 在一些实施例中,聚合物可W根据本发明进行使用而不经历交联步骤。 In some embodiments, the polymer may be subjected to W without crosslinking step according to the present invention is used. 进一步理解的是,本发明的合成纳米载体可W包含嵌段共聚物、接枝共聚物、共混物、混合物、和/或任何上述及其他聚合物的加合物。 Further it understood that the synthetic nanocarriers of the present invention may comprise W block copolymers, graft copolymers, blends, mixtures, and / or any other adduct and the above-described polymers. 本领域的那些技术人员将认识到,在此列出的运些聚合物代表根据本发明可W使用的聚合物的示例性的、而不是全面的清单。 Those skilled in the art will recognize that the transport of these polymers represent listed here, not a comprehensive list of exemplary polymers according to the present invention W may be used.

[0169] 根据本发明的多种组合物包含与药学上可接受的赋形剂(如防腐剂、缓冲剂、盐水或憐酸盐缓冲盐水)组合的合成纳米载体。 [0169] According to various combinations of the compositions of the present invention comprise a pharmaceutically acceptable excipient (such as preservatives, buffers, saline, or saline buffered salt pity) synthetic nanocarriers. 可W使用常规药物制造和配制技术制造运些组合物,W实现有用的剂型。 W may be manufactured using conventional pharmaceutical formulating techniques and manufacturing operation these compositions, W useful dosage forms. 在一个实施例中,将本发明的合成纳米载体与防腐剂一起悬浮在注射用无菌盐水溶液中。 In one embodiment, suspended in sterile saline injectable aqueous medium together with a preservative synthetic nanocarriers of the present invention.

[0170] 在多个实施例中,当制备合成纳米载体作为载体时,用于将组分与合成纳米载体偶联的方法可W是有用的。 [0170] In various embodiments, when preparing synthetic nanocarriers as a carrier, a method for the coupling component to the synthetic nanocarriers W may be useful. 如果该组分是小分子,那么在组装运些合成纳米载体之前将该组分附接至聚合物上可能是有利的。 If the component is a small molecule, then the set before shipment of these synthetic nanocarriers component attached to the polymer may be advantageous. 在多个实施例中,制备具有用于将该组分与合成纳米载体偶联的表面基团的合成纳米载体也可能是有利的,该偶联是通过使用运些表面基团而不是将该组分附接至聚合物上且随后在合成纳米载体的构建中使用运种聚合物共辆物来进行。 In various embodiments, for the preparation of a synthetic nanocarriers surface groups of the component with the synthetic nanocarriers coupling may also be advantageous to transport these coupling by using instead of the surface groups component attached to the polymer, and subsequently in the co-transport vehicles polymers was constructed using synthetic nanocarriers.

[0171] 在某些实施例中,该偶联可W是共价连接物。 [0171] In certain embodiments, the coupling can be W is a covalent linker. 在多个实施例中,根据本发明的肤可W经由1,2,3-Ξ挫连接物共价偶联至外表面上,该连接物通过纳米载体表面上的叠氮基团与含有烘基的抗原或免疫抑制剂进行1,3-偶极环加成反应、或通过纳米载体表面上的烘与含有叠氮基团的组分进行1,3-偶极环加成反应而形成。 In various embodiments, the coupling via frustrated 1,2,3-Ξ covalently to the outer surface of the skin according to the present invention may be W, the linker group by azide on the surface of the nanocarrier containing drying group antigen or immunosuppressant 1,3-dipolar cycloaddition reaction, or are formed by baking on the surface of the nanocarrier containing the component stack nitrogen groups the 1,3-dipolar cycloaddition reaction. 此类环加成反应优选地是在铜(I) 催化剂W及适合的Cu(I)-配体和将Cu(II)化合物还原成催化活性的Cu(I)化合物的还原剂存在下进行的。 Such a cycloaddition reaction is preferably copper (I) catalyst and a suitable W Cu (I) - for the reduction and ligand (II), Cu as the reducing agent in the presence of compound (I) under the catalytic activity of Cu . 此类Cu(I)催化的叠氮-烘环加成反应(CuAAC)也可W称为点击反应。 Such Cu (I) catalyzed azide - drying cycloaddition reaction (CuAAC) W may also be referred to as a click reaction.

[0172] 另外,共价偶联可W包含共价连接物,该共价连接物包含酷胺连接物、二硫化物连接物、硫酸连接物、腺连接物、酷阱连接物、亚胺或朽连接物、脈或硫脈连接物、脉连接物、胺连接物、W及横酷胺连接物。 [0172] Further, covalent coupling may comprise a covalent linker W, the covalent linkage comprises a Cool amine linker, a disulfide linker, connected to a sulfate, glandular linker, linker cool trap, imine or rotten linker, or a thiourea linker pulse, pulse linker, an amine linker, W, and an amine cross-linker cool.

[0173] 酷胺连接物经由一种组分上的胺与第二组分(如纳米载体)的簇酸基团之间的酷胺键形成。 [0173] Cool cool via an amine linker amine linkage is formed between the amine and the second component (such as a nanocarrier) on one component of the cluster acid groups. 该连接物中的酷胺键可W用被适当保护的氨基酸或组分与被活化的簇酸(如被N-径基下二酷亚胺活化的醋)、使用任何常规酷胺键形成反应制成。 The linker may be the cool amine linkage W (such as are activated under two imine cool-yl acetate N- diameter) with a suitably protected amino acids or components of the cluster acid and activated using any conventional cool amine bond formation production.

[0174] 二硫化物连接物经由在例如R1-SS-R2的形式的两个硫原子之间形成二硫(SS) 键制成。 [0174] via a disulfide linker form a disulfide (SS) between two sulfur atoms in the form of, for example, R1-SS-R2 bond is made. 二硫键可W通过含有硫醇基/琉基(-SH)的组分与聚合物或纳米载体上另一个被活化的硫醇基或含有硫醇基/琉基的纳米载体与含有被活化的硫醇基的组分进行硫醇交换形成。 W may be a disulfide bond by the polymer component containing a thiol group or a nanocarrier / thiol (-SH) of the other activated thiol group or thiol group containing nanocarrier / sulfur containing group is activated component thiol group of a thiol exchange form. 村一1·^. Village · 1 ^.

[01巧]Ξ挫连接物(特别是其中R1和R2可W是任何化学实体的形式^的1,2,3-Ξ挫) 朽3. 是通过附接至第一组分(如纳米载体)上的叠氮基与附接至第二组分(如免疫抑制剂或抗原)上的末端烘的1,3-偶极环加成反应制成。 [Qiao 01] a Cascade frustrated linker (particularly wherein W is R1 and R2 may be any chemical entity in the form of 1,2,3-Ξ ^ setback) 3. rotten by attached to the first component (e.g., the nanocarrier formed on the end of drying the 1,3-dipolar azide group is attached to the second component (e.g., an immunosuppressant or antigen) on a) cycloaddition reaction. 该1,3-偶极环加成反应在有或无催化剂的情况下、优选地在有经由1,2,3-Ξ挫官能团连接两个组分的Cu(I)催化剂的情况下进行。 The 1,3-dipolar cycloaddition reaction in the presence or absence of a catalyst, preferably in the case where there is performed fell functional groups of the two components Cu (I) catalyst via a 1,2,3-Ξ. 夏普里斯(Sharpless)等人,在德国应用化学(AngewXhem. Int.Ed.) ,41(14) ,2596,(2002)中W 及梅尔达尔(Meldal)等人,在化学评述(Chem.Rev. ),2008,108(8) ,2952-3015中详细描述了此化学作用,并且该化学作用常常被称为"点击"反应或CuAAC。 Sharp Rees (Sharpless) and others, used in the German chemical (AngewXhem. Int.Ed.), 41 (14), 2596, (2002) in W and Mel Dahl (Meldal) and others, in chemically comments (Chem.Rev .), 2008,108 (8), 2952-3015 described in detail in this chemistry, and the chemistry is often referred to as "click" or CuAAC reaction.

[0176]在多个实施例中,制备了一种在聚合物链末端含有一个叠氮基或烘基的聚合物。 [0176] In various embodiments, the polymer containing an azido group or baked in the polymer chain ends prepared. 然后使用此聚合物来制备合成纳米载体,W此方式使得多个烘或叠氮基放置在该纳米载体的表面上。 Is then prepared using the polymer synthesized nanocarriers, W in this way that a plurality of drying or azido placed on the surface of the nanocarrier. 可替代地,可W通过另一种途径制备该合成纳米载体,并且随后用烘或叠氮基功能化。 Alternatively, W may be prepared by the synthetic nanocarriers another way, and then by drying or azido functional. 该组分是在或者烘(如果该聚合物含有叠氮基)或者叠氮基(如果该聚合物含有烘)存在下制备。 This component is baked or (if the polymer containing an azido) or azido (if the polymer contains baking) prepared in the presence. 然后在具有或不具有催化剂的情况下允许该组分与该纳米载体经由1,3-偶极环加成反应进行反应,该催化剂将该组分经由1,4-二取代的1,2,3-Ξ挫连接物共价偶联至颗粒上。 Then allowed in the case with or without a catalyst component is reacted via the 1,3-dipolar cycloaddition reaction with the nanocarrier, the catalyst component the 1,2-substituted 1,4-through, 3-Ξ setback linker is covalently coupled to the particles.

[0177] 硫酸连接物通过W例如R1-S-R2的形式形成硫-碳(硫酸)键制成。 [0177] connected to form sulfur by a sulfated form of, for example, W R1-S-R2 is - C (acid) key is made. 硫酸可W通过用第二组分(如纳米载体)上的烷基化基团(如面基或环氧基)烷基化一种组分(如该组分)上的硫醇基/琉基(-SH)制成。 W sulfuric acid can be alkylated by treatment with a second component group (e.g., epoxy group or plane) on (e.g. nanocarriers) alkylation of one component (e.g., the component) thiol / sulfur on group (-SH) is made. 硫酸连接物还可W通过将一种组分上的硫醇基/琉基与作为迈克尔受体的含有马来酷亚胺基团或乙締讽基团的第二组分(如一种聚合物)上的缺电子締基进行迈克尔加成来形成。 W may also be connected by a sulfate group on one component of a thiol / thiol groups to maleic cool imino or the second component B Bitterness associative group (e.g., a polymer containing as Michael acceptor associating the electron deficient group) to form a Michael addition. W另一种方式,硫酸连接物可W通过一种组分上的硫醇基/琉基与第二组分(如一种聚合物或纳米载体)上的締基进行自由基硫醇-締反应来制备。 Another embodiment W, W may be connected to a sulfate radical through a thiol group on one component of the thiol / thiol and a second component (e.g., a polymer or a nanocarrier) based on the association - association reaction be prepared.

[0178] 腺连接物通过一种组分上的酷阱基与第二组分(如纳米载体)上的醒基/酬基进行反应制成。 [0178] gland connection was made by reacting cool wellbase second component (e.g., the nanocarrier) wake group / groups paid on the one component.

[0179] 酷阱连接物通过一种组分上的阱基与第二组分(如纳米载体)上的簇酸基团进行反应来形成。 [0179] Cool well linker formed by reacting the acid groups on the cluster well groups with a second component (such as a nanocarrier) on one component. 此类反应通常使用类似于形成酷胺键的化学方法(其中簇酸用活化试剂活化) 来进行。 Such reactions are typically formed using chemical methods analogous to cool amine linkage (where the cluster acid activation with an activating reagent) is performed.

[0180] 亚胺或目亏连接物通过一种组分上的胺或N-烷氧基胺基(或氨氧基)与第二组分(如纳米载体)上的醒或酬基进行反应来形成。 [0180] imine or head loss linker amine or by reaction of N- alkoxy group (or amino group) on one component and a second component (such as a nanocarrier) group on waking or reward is formed.

[0181] 脈或硫脈连接物通过一种组分上的胺基与第二组分(纳米载体)上的异氯酸醋或异硫氯酸醋基进行反应来制备。 [0181] veins or thiourea linker prepared by reacting an amine with a second component (nanocarriers) isocyanurate acid vinegar or acetic acid isothiocyanato groups on the one component.

[0182] 脉连接物通过一种组分上的胺基与第二组分(如纳米载体)上的亚氨酸醋基团进行反应来制备。 [0182] Pulse linkers prepared by amine groups and a second component (such as a nanocarrier) alkylene acetic acid group on one component on the reaction.

[0183] 胺连接物通过一种组分上的胺基与第二组分(纳米载体)上的烷基化基团(如面基、环氧基、或横酸醋基)进行烷基化反应制成。 [0183] alkylated amine linker group by alkylation (e.g., the surface group, an epoxy group, or a cross-yl vinegar) on the amine groups and a second component (nano-carriers) on one component The reaction is made. 可替代地,胺连接物还可W用适合的还原试剂(如氯基棚氨化钢或Ξ乙酷氧基棚氨化钢)、通过一种组分上的胺基与第二组分(如纳米载体)上的醒基或酬基进行还原胺化反应制成。 Alternatively, the amine linker W also with a suitable reducing agent (e.g., steel or amide-chloro-shed B Ξ cool shed amide group steel), the second component via amine groups on one component ( the wake-yl group or reward on a nanocarrier) is made of a reductive amination reaction.

[0184] 横酷胺连接物通过一种组分上的胺基与第二组分(纳米载体)上的横酷基面化物(如横酷氯)基团进行反应制成。 [0184] Cool a cross-linker via an amine group and a second component (nano-carriers) on the surface of a component of a cross-yl cool on the compound (e.g., cross-chloro cool) made of a group for the reaction.

[0185]讽连接物通过亲核试剂与乙締讽进行迈克尔加成制成。 [0185] Bitterness connection was made by Michael addition of nucleophiles associated with B ridicule. 或者该乙締讽或者该亲核试剂可W位于纳米载体的表面上或附接至一种组分上。 Bitterness associated or on the B or W may be the nucleophile of the nanocarrier, or attached to the surface of one component.

[0186] 该组分还可W经由非共价共辆方法与纳米载体共辆。 [0186] The W component may vehicle via a non-covalent manner were co nanocarrier vehicles. 例如,带负电荷的抗原或免疫抑制剂可W通过静电吸附与带正电荷的纳米载体共辆。 For example, negatively charged antigen or immunosuppressant W may be formed by electrostatically attracting the positively charged carrier nano co vehicle. 含有金属配体的组分还可W经由金属-配体络合物与含有金属络合物的纳米载体共辆。 Ligand containing a metal component may also W via the metal - ligand complex with a metal complex containing a nanocarrier total units.

[0187] 在多个实施例中,该组分可W在组装该合成纳米载体之前附接至聚合物(例如聚乳酸-嵌段-聚乙二醇)上,或可W形成该合成纳米载体而使得反应性或可活化基团在它的表面上。 [0187] In various embodiments, the components can be assembled before the W nanocarrier attached to synthetic polymers (e.g. polylactic acid - block - polyethylene glycol) on, or forming the synthetic nanocarriers W so that reactive or activatable groups on its surface. 在后一种情况下,可W制备该组分W使其具有与由运些合成纳米载体的表面呈现的附接化学性质相容的基团。 In the latter case, the component may be prepared W W group it has compatibility with the chemical attachment properties exhibited by the surface of the synthetic nanocarriers some transport. 在其他实施例中,可W使用一种适合的连接物将肤组分附接至化P或脂质体上。 In other embodiments, W may be the use of a suitable linker is attached to the skin of the component P or liposomes. 连接物是能够将两个分子连接在一起的化合物或试剂。 It is a linker capable of linking together two molecules of the compound or agent. 在一个实施例中,该连接物可W是在赫曼森化ermanson)2008中所描述的同双功能或异双功能试剂。 In one embodiment, the linker can be of W in Hermanson ermanson) homobifunctional or 2008 described heterobifunctional reagent. 例如,可W在抓C存在下用同双功能连接物(己二酸二酷阱(ADH))处理表面上含有簇基的化P 或脂质体合成纳米载体,W形成具有ADH连接物的相应合成纳米载体。 For example, W can be arrested in the presence of C was treated with homobifunctional linker (adipic cool trap (ADH)) of the cluster group P or liposomes containing synthetic nanocarriers on the surface, forming a W ADH linker corresponding synthetic nanocarriers. 然后使所得被ADH连接的合成纳米载体经由NC上的该ADH连接物的另一端与含有酸基团的肤组分共辆,W产生相应的VLP或脂质体肤共辆物。 The resulting synthetic nanocarriers then connected via the other end of the ADH ADH linker on the skin NC component containing an acid group-containing conjugated units, W generates a corresponding VLP or liposome vehicles was co skin.

[0188] 针对可获得的共辆方法的详细描述,参见2008年由学术出版公司(Academic Press, Inc.)出版的第2 版,赫曼森《GT(He;rmanson G Τ)。 [0188] for a detailed description of the vehicles were available methods, see published in 2008 by the Academic Publishing Company (Academic Press, Inc.) version 2, Hermanson "GT (He; rmanson G Τ). 生物共辆技术(Bioconjugate Techniques)"。除了共价附接之外,该组分可W通过吸附至预先形成的合成纳米载体上来偶联、或它可W通过在该合成纳米载体形成过程中进行封装来偶联。 Organism vehicles art (Bioconjugate Techniques) ". In addition covalently attached outside, this component can be coupled by adsorption W up to a pre-formed synthetic nanocarriers, or it may be W by the synthesis process for forming nanocarrier encapsulation coupling.

[0189] 在此所提供的任何免疫抑制剂可W与该合成纳米载体偶联。 [0189] Any immunosuppressive agents provided herein may be conjugated to the vector W Nanometer. 免疫抑制剂包括但不限于:抑制素;mTOR抑制剂,如雷帕霉素或一种雷帕霉素类似物;TGF-β信号剂;TGF-β受体激动剂;组蛋白去乙酷化酶化DAC)抑制剂;皮质类固醇类;线粒体功能抑制剂,如鱼藤酬;P38 抑制剂;NF-κβ抑制剂;腺巧受体激动剂;前列腺素E2激动剂;憐酸二醋酶抑制剂,如憐酸二醋酶4抑制剂;蛋白酶体抑制剂;激酶抑制剂;G-蛋白偶联受体激动剂;G-蛋白偶联受体括抗剂;糖皮质激素类;类视黄醇类;细胞因子抑制剂;细胞因子受体抑制剂;细胞因子受体活化剂;过氧化物酶体增殖物激活受体括抗剂;过氧化物酶体增殖物激活受体激动剂;组蛋白去乙酷化酶抑制剂;巧调憐酸酶抑制剂;憐酸酶抑制剂W及氧化的ATP。 Immunosuppressive agents include but are not limited to: inhibin; of mTOR inhibitors such as rapamycin or one rapamycin analogues; TGF-β signal reagent; TGF-β receptor agonists; histone cool of B enzymatic DAC) inhibitors; corticosteroids; mitochondrial function inhibitors such as rotenone pay; inhibitor of P38; NF-κβ inhibitor; Qiao adenovirus receptor agonist; prostaglandin E2 agonist; pity acid esterase inhibition agent, such as 4 pity acid esterase inhibitors; proteasome inhibitors; kinase inhibitors; G-protein coupled receptor agonists; G-protein coupled receptor antagonist comprising; glucocorticoids; retinoid alcohols; cytokine inhibitors; inhibitors of cytokine receptors; cytokine receptor activator; peroxisome proliferator-activated receptor antagonist comprising; peroxisome proliferator-activated receptor agonist; group cool to b protein inhibitors; clever modulation pity acid inhibitor; pity acid oxidation inhibitor and W ATP. 免疫抑制剂还包括ID0、维生素D3、环抱霉素A、芳香控受体抑制剂、白襲芦醇、硫挫嚷岭、6-琉基嚷岭、阿司匹林、尼氣酸、雌立醇、雷公藤甲素、白细胞介素(例如,比-1、比-1〇)、环抱霉素A、siRNA祀向细胞因子或细胞因子受体等。 Further comprising the ID0 immunosuppressant, vitamin D3, cyclosporine A, control aromatase inhibitor receptor, white passage resveratrol, sulfur setback cried Ridge, 6-thiol cried Ridge, aspirin, Nigeria acid gas, alcohol female Li, Thunder Triptolide, interleukins (e.g., ratio -1 -1〇 ratio), cyclosporine A, siRNA to worship cytokine or cytokine receptor and the like.

[0190] 抑制素的实例包括:阿托伐他汀(atorvastatin)( UPn.'OR-\TORVAS'r* )v 西立伐他汀(cerivastatin)、氣伐他汀(f luvastatin) (LESC0.L*>、LESCOL'。'化)、洛伐他汀(1 ο V astati η ) ( mevacOR气AUrOCOR" wVLTOPREVa )、美伐他汀(mevastatin) (COMMCTIN;® )、匹伐他汀(Pitavastatin) (IJVAIX):*、pjaVA® )、罗苏伐他汀(的311乂日3*日^11)(1>化4\;\(:〇01/、SELEKTINE -、LIPOSTAT') 罗苏伐他汀(CRESTOR*似及辛伐他汀(simvastatin) (ZOCO民UPEX* )。 Examples [0190] statins include: atorvastatin (atorvastatin) (UPn.'OR- \ TORVAS'r *) v statin cerivastatin (cerivastatin), fluvastatin gas (f luvastatin) (LESC0.L *> '.', LESCOL of), lovastatin (1 ο V astati η) (mevacOR gas AUrOCOR "wVLTOPREVa), mevastatin (mevastatin) (COMMCTIN; ®), pitavastatin (pitavastatin) (IJVAIX): *, pjaVA®), rosuvastatin (311 ^ qe day 3 * day 11) (1> of 4 \; \ (: 〇01 /, SELEKTINE -, LIPOSTAT ') rosuvastatin (simvastatin and the like of CRESTOR * statins (simvastatin) (ZOCO people UPEX *).

[0191] mTOR抑制剂的实例包括:雷帕霉素和其类似物(例如,C化-779、RAD001、AP23573、 C20-甲代締丙基雷帕霉素(C20-Marap)、C16-(S)-下基横酷氨基雷帕霉素(C16-BSrap)、 C16-(S)-3-甲基吗I噪雷帕霉素(C16-iRap)(贝尔(Bayle)等人,化学与生物学(Chemis1:ry& Biology)2006,13:99-107))、AZD8055、BEZ235(NVP-BEZ235)、大黄根酸(大黄酪)、德福罗莫司(deforolimus) (MK-8669)、依维莫司(everolimus) (RAD0001)、KU-0063794、PI_103、 PP242、替西罗莫司(temsirolimus) W及WYE-354(可获自赛立克公司(Selleck),休斯顿化ouston),德克萨斯州(TX),美国(USA))。 Examples [0191] mTOR inhibitors include: rapamycin and its analogs (e.g., C of -779, RAD001, AP23573, C20- methallyl propionate association rapamycin (C20-Marap), C16- ( S) - cool the ammonia-based cross-rapamycin (C16-BSrap), C16- (S) -3- methylmorpholine I noise rapamycin (C16-iRap) (Bell (Bayle) et al., Chemistry and Biology (Chemis1: ry & Biology) 2006,13: 99-107)), AZD8055, BEZ235 (NVP-BEZ235), rhubarb root acids (casein rhubarb), Telford lomustine (deforolimus) (MK-8669), by everolimus (everolimus) (RAD0001), KU-0063794, PI_103, PP242, temsirolimus (temsirolimus) W and WYE-354 (available from Rickettsia game company (Selleck), of Houston ouston), Texas Texas (TX), United states of America (USA)).

[0192] TGF-β信号剂的实例包括:TGF-β配体(例如,活化素4、60。1、60。11、骨形态发生蛋白、noda 1、TGF-β)和它们的受体(例如,ACVR1B、ACVR1C、ACVR2A、ACVR2B、BMPR2、BMPR1A、 BMPR1B、TG邱RI、TG邱RII)、R-SMADS/co-SMADS(例如,SMAD1、SMAD2、SMAD3、SMAD4、SMAD5、 SMAD8)、W及配体抑制剂(例如,卵泡抑素、头蛋白、腫蛋白、DAN、lefty、LTBPl、T皿Sl、核屯、 蛋白聚糖)。 [0192] Examples of TGF-β signal include: TGF-β ligand (e.g., 4,60.1,60.11 activin, bone morphogenic proteins, noda 1, TGF-β) and their receptors ( For example, ACVR1B, ACVR1C, ACVR2A, ACVR2B, BMPR2, BMPR1A, BMPR1B, TG Qiu RI, TG Qiu RII), R-SMADS / co-SMADS (e.g., SMAD1, SMAD2, SMAD3, SMAD4, SMAD5, SMAD8), W and ligand inhibitors (e.g., follistatin, noggin, swollen protein, DAN, lefty, LTBPl, T dish Sl is, nuclear Tun, proteoglycans).

[0193] 线粒体功能抑制剂的实例包括:苍术巧(二钟盐)、米酵菌酸(Ξ锭盐)、幾基氯化间氯苯腺、簇基苍术巧(例如,来自苍术属植物(Atractylis gummifera))、CGP-37157、(-)-鱼藤素(例如,来自栓皮豆(Mundulea sericea))、F16、己糖激酶II VDAC结合域肤、寡霉素、鱼藤酬、Ru360、SFKl、W及鄉氨霉素(例如,来自极暗黄链霉菌(Streptomyces fulvissimus)) 化MD4生物科学化MD4Biosciences),美国化SA))。 [0193] Examples of inhibitors of mitochondrial function comprising: herb Qiao (two clock salt), BKA (a Cascade ingot salt), among several trichloride chlorophenyl gland, herb clever cluster group (e.g., derived from the herb genus ( Atractylis gummifera)), CGP-37157, (-) - deguelin (e.g., from cork beans (Mundulea sericea)), F16, hexokinase II VDAC binding domain skin, oligomycin, rotenone paid, Ru360, SFKl , W and township ammonia neomycin (eg, from a very dark yellow Streptomyces (Streptomyces fulvissimus)) of MD4 biological scientific MD4Biosciences), United States of SA)).

[0194] P38抑制剂的实例包括:SB-203580(4-(4-氣苯基)-2-(4-甲基亚硫酷基苯基)-5- (4-化晚基)1H-咪挫)、SB-239063(反式-1-(4-¾基环己基)-4-(氣苯基)-5-(2-甲氧基-喀晚-4-基)咪挫)、SB-220025(5-(2-氨基-4-喀晚基)-4-(4-氣苯基)-1-(4-赃晚基)咪挫))、 W及ARRY-797。 Example [0194] P38 inhibitors include: SB-203580 (4- (4- gas phenyl) -2- (4-methylsulfinyl phenyl cool-yl) -5- (4-LATE-yl) lH- imidazol setback), SB-239063 (trans -1- (4-¾ cyclohexyl) -4- (gas) -5- (2-methoxy - Ka Night 4-yl) imidazole setback), SB-220025 (5- (2- amino-4-yl Night Cameroon) -4- (4-gas-phenyl) -1- (4-stolen Night-yl) imidazole setback)), W, and ARRY-797.

[0195] NF(例如,ΝΚ-κβ)抑制剂的实例包括;IF畑1、2-(1,8-糞晚-2-基)-苯酪、5-氨基水杨酸、BAY 11-7082、BAY 11-7085、〔4阳(咖啡酸苯乙醋)、马来酸二乙醋、1邸-2抑制剂1¥、 IMD 0354、乳胞素(lac化巧stin)、MG-132口-Leu-Leu-Leu-C册]、NFkB活化抑制剂III、NF-K B活化抑制剂II、JSH-23、小白菊内醋(partheno 1 ide)、氧化苯肿(PA0)、PPM-18、化咯烧二硫代氨基甲酸锭盐、QNZ、R0 106-9920、棟酷胺(rocaglamide)、棟酷胺AL、棟酷胺C、棟酷胺I、 棟酷胺J、洛克米兰醇(rocaglaol)、(R)-MG-132、水杨酸钢、雷公藤甲素(PG490)、膨峡菊内醋(wedelolactone)。 [0195] NF (e.g., ΝΚ-κβ) inhibitors include; Hata the IF 1,2- (1,8 fecal Night 2-yl) - benzene casein, 5-aminosalicylic acid, BAY 11-7082 , BAY 11-7085, [4 male (caffeic acid phenethyl acetate), maleic acid ethyl ester, 1 dI-2 inhibitor 1 ¥, IMD 0354, lactacystin (lac of clever stin), MG-132 port -Leu-Leu-Leu-C books], NFkB activation inhibitor III, NF-K B activation inhibitor II, JSH-23, the feverfew vinegar (partheno 1 ide), oxidation of benzene swelling (PA0), PPM-18 , slightly burning of the ingot dithiocarbamate salts, QNZ, R0 106-9920, Dong cool amine (rocaglamide), cool buildings amine AL, Dong C cool amines, amine cool buildings I, J amine cool buildings, Locke Milan alcohol ( rocaglaol), (R) -MG-132, salicylic steel, triptolide (PG490), Ju-expanding Gap vinegar (wedelolactone).

[0196] 腺巧受体激动剂的实例包括CGS-21680和ATl-146e。 [0196] Examples of coincidence adenovirus receptor agonists include CGS-21680 and ATl-146e.

[0197] 前列腺素E2激动剂的实例包括E-前列腺素类激素2化-Prostanoid 2)和E-前列腺素类激素4。 [0197] Examples of prostaglandin E2 agonists include E- prostanoids 2 of -Prostanoid 2) and 4 E- prostanoids.

[0198] 憐酸二醋酶抑制剂(非选择性和选择性抑制剂)的实例包括:咖啡因、氨茶碱、IBMX (3-异下基-1-甲基黄嚷岭)、副黄嚷岭、己酬可可碱、可可碱、茶碱、甲基化的黄嚷岭类、长春西汀(vi吨ocetine)、EHNA(赤-9-(2-¾基-3-壬基)腺嚷岭)、阿那格雷(anagrelide)、伊诺昔酬(enoximone) (PERFAN™)、米利酬(milrinone)、左昔孟坦(levosimendon)、松叶菊碱(mesembrine)、异下司特(ibudilast)、R比拉米司特(piclamilast)、木犀草素(luteolin)、 屈他维林(drotaverine)、罗氣司特(roflumilast) (DAXAS™、DALIRESP™)、西地那非(sildenafil)( relation。'、VIAGRA:® )、他达手立非(tadalafil)( ADO权CA®'、CIAL扮*)、伐地那非(vardenafil)(LEVITRA®、SmXYNii')、乌地那非(udenafil)、阿伐那非(avanafil)、淫羊蕾巧(icariin)、4-甲基赃嗦、W及R比挫并喀晚-7- 1〇 [0198] Examples of pity acid esterase inhibitor (selective and non-selective inhibitors) include: (yellow-1-methyl-3-isobutyl cried the ridge), yellow sub caffeine, theophylline, IBMX cried Ridge, hexyl pay theobromine, theobromine, theophylline, methylated yellow cried ridge type, vinpocetine (vi t ocetine), EHNA (erythro -9- (2-¾-3-nonyl) gland cried Ridge), anagrelide (anagrelide), Eno past paid (enoximone) (PERFAN ™), Millie pay (milrinone), left Xi Meng Tan (levosimendon), Mesembryanthemum base (mesembrine), under different Sterling (ibudilast) , R Bi Lami Sterling (piclamilast), luteolin (luteolin), drotaverine (drotaverine), Lo gas zafirlukast (roflumilast) (DAXAS ™, DALIRESP ™), sildenafil (sildenafil) (relation . ', VIAGRA: ®), he reached a hand stand non (tadalafil) (ADO rights CA®', CIAL play *), vardenafil (vardenafil) (LEVITRA®, SmXYNii '), Udenafil (udenafil) , avanaphil (avanafil), epimedium bud Qiao (icariin), 4- methyl winded stolen, W, and R ratio Ka and night setback -7- 1〇

[0199] 蛋白酶体抑制剂的实例包括棚替佐米(bortezomib)、双硫仑(disulf iram)、表儿茶素-3-没食子酸盐、W及盐抱菌酷胺A(salinosporamide A)。 [0199] Examples of proteasome inhibitors include bortezomib shed (bortezomib), disulfiram (disulf iram), epicatechin gallate -3-, W, and amine salts hold bacteria cool A (salinosporamide A).

[0200] 激酶抑制剂的实例包括:贝伐单抗(bevacizumab)、BIBW 2992、西妥昔单抗(cetuximabH ER技1TUX® )、伊马替尼(imatinibH glee VE(:@ )、曲妥单抗(trastuzumab) ( HERCEFON*)、吉非替尼(gef itinib) (I说祕)、兰尼单抗(ranibizumab) (LU-CENT.蚁!《)、赃加他尼(pegap1:anib)、索拉非尼(sorafen;Lb)、达沙替尼(dasatin;Lb)、舒尼替尼(sunitin;Lb)、埃罗替尼(e;rlotin;Lb)、尼罗替尼(nilotinib)、拉帕替尼(lapatinib)、帕尼单抗(panitumumab)、凡德他尼(vandetanib)、E7080、帕挫帕尼(pazopanib)、木矛U替尼(mubritinib)。 Examples [0200] kinase inhibitors include: Avastin (bevacizumab), BIBW 2992, cetuximab (cetuximabH ER technology 1TUX®), imatinib (imatinibH glee VE (: @), trastuzumab (.! LU-CENT ants ") anti- (trastuzumab) (HERCEFON *), gefitinib (gef itinib) (I say secret), ranibizumab (ranibizumab), stolen pegaptanib (pegap1: anib), sorafenib (sorafen; Lb), dasatinib (dasatin; Lb), sunitinib (sunitin; Lb), erlotinib (e; rlotin; Lb), nilotinib (nilotinib), lapatinib (lapatinib), panitumumab (panitumumab), vandetanib (vandetanib), E7080, Pa frustrated Trapani (pazopanib), wooden spears U erlotinib (mubritinib).

[0201] 糖皮质激素类的实例包括:氨化可的松(皮质醇)、醋酸可的松、强的松(prednisone)、泼尼松龙(predn iso lone)、甲泼尼龙(me thylpredn iso lone)、地塞米松(dexamethasone)、倍他米松(betamethasone)、曲安西龙(triamcinolone)、倍氯米松(beclometasone)、醋酸氣氨可的松、醋酸脱氧皮质酬(D0CA)、W及醒固酬。 [0201] Examples of glucocorticoids include: amide cortisone (cortisol), cortisone acetate, prednisone (prednisone), prednisolone (predn iso lone), methylprednisolone (me thylpredn iso Lone), dexamethasone (dexamethasone), betamethasone (betamethasone), triamcinolone (triamcinolone), beclomethasone (beclometasone), ammonia gas cortisone acetate, deoxycorticosterone acetate paid (D0CA), W and wake solid pay.

[020^ 类视黄醇类的实例包括:视黄醇、视黄醒、维甲酸(视黄酸,防ΕΤΪΝ-Α'α0、异维甲酸(Accutiine®、Anmesteein@、Oaravis汲、Sotret®)、阿利维甲酸(曰1 i tret ino iη)( PA;NRETIN@ )、依曲替醋(e 化etinate)(TEGI SONT™ )和它的代谢物阿维A (ac i tr eti η)( SORIATANE…)、他扎罗汀(tazarotene) (Tazoim'"、A、'age"、Zorac® )、贝沙罗汀(bexarotene) (TARGRETIN6 )、W及阿达帕林(adapalene) (DIFFERIN'6;)。 [Example 020 ^ retinoids include: retinol, retinyl awake, tretinoin (retinoic acid, anti ΕΤΪΝ-Α'α0, isotretinoin (Accutiine®, Anmesteein @, Oaravis drain, Sotret®) , Allie retinoic acid (say 1 i tret ino iη) (PA; NRETIN @), according to music for vinegar (e of etinate) (TEGI SONT ™) and its metabolite acitretin a (ac i tr eti η) (SORIATANE ...), tazarotene (tazarotene) (Tazoim ' ", A,' age", Zorac®), bexarotene (bexarotene) (TARGRETIN6), W and adapalene (adapalene) (DIFFERIN'6;) .

[0203] 细胞因子抑制剂的实例包括ILlra、ILl受体括抗剂、IGFBP、TNF-BF、尿调节素(uromodulin)、a-2 -巨球蛋白、环抱霉素A、戊烧脉、W及己酬可可碱( Pentopak;气Peirto础®v TrentafO。 [0203] Examples of cytokine inhibitors include ILlra, ILl receptor antagonist including, IGFBP, TNF-BF, uromodulin (uromodulin), a-2 - macroglobulin, cyclosporine A, pentyl burning pulse, W and have paid theobromine (Pentopak; gas Peirto basis ®v TrentafO.

[0204] 过氧化物酶体增殖物激活受体括抗剂的实例包括GW9662、PPAR 丫括抗剂III、 G335、T0070907 化MD4 生物科学化MD4Biosciences),美国化SA))。 [0204] Peroxisome proliferator-activated receptor antagonist comprising Examples include GW9662, PPAR antagonist including Ah III, G335, T0070907 of MD4 biological scientific MD4Biosciences), United States of SA)).

[0205] 过氧化物酶体增殖物激活受体激动剂的实例包括:化格列酬(pioglitazone)、环格列酬レigli化zone)、氯贝特レlofibrate)、GW1929、GW7647、レ165,041、LY 171883、PPAR 丫活化剂、Fmoc-Leu、曲格列酬(troglitazone)、W 及WY-14643(EMD4生物科学化MD4Biosciences),美国化SA))。 [0205] Examples of peroxisome proliferator-activated receptor agonists include: cell columns of pay (pioglitazone), Rayon ring igli of glibenclamide pay zone), clofibrate Rayon lofibrate), GW1929, GW7647, Rayon 165,041 , LY 171883, PPAR activator Ah, Fmoc-Leu, troglitazone pay (troglitazone), W, and WY-14643 (EMD4 biological scientific MD4Biosciences), United States of SA)).

[0206] 组蛋白去乙酷化酶抑制剂的实例包括:异氨朽酸类(或异氨朽酸醋类),如曲古抑菌素AUrichostatin A);环状四肤类(如曲破辛B(trapoxin B))和缩肤类;苯甲酯胺类;亲电酬类;脂肪族酸化合物,如苯基下酸醋和丙戊酸;异径朽酸类,如伏立诺他(vorinos化t) (SAHA)、贝林司他(belinostat)( PXD101)、LAQ824、W及帕比司他(panobinostat) (LBH589);苯甲酯胺类,如恩替诺特(entinostat)(MS-275)、CI994、W及莫替司他(mocetinostat) (MGCD0103)、烟酷胺;NAD的衍生物、二氨香豆素、糞并R比喃酬(naphthopyranone)、W及2-径基糞醒类。 [0206] Examples of group B proteins to cool inhibitors include: iso-amino acids rot (iso ammonia or rotten Suancu class), such as trichostatin AUrichostatin A); cyclic tetrapeptides skin (such as broken curve oct-B (trapoxin B)) and reduced skin type; benzyl ester amines; electrophilic pay the like; aliphatic acid compounds, such as valproic acid and vinegar under phenyl; rot reducers acids, such as vorinostat (vorinos of t) (SAHA), Belinostat (belinostat) (PXD101), LAQ824, W and Panobinostat (panobinostat) (LBH589); phenylmethyl ester amines, such entecavir Knott (entinostat) ( MS-275), CI994, W and Mo for orlistat (mocetinostat) (MGCD0103), cool smoke amine; the NAD derivative, coumarin diamino, unpaid thiopyran manure and R ratio (naphthopyranone), W-diameter and 2 manure wake base class.

[0207] 巧调憐酸酶抑制剂的实例包括环抱霉素、化美莫司(pimecrolimus)、伏环抱素(voclosporin)、W及他克莫司(tacrolimus)。 Examples [0207] Qiao tone Rei acid inhibitors include cyclosporine, Grace Division Mo (pimecrolimus), V cyclosporine (voclosporin), W and tacrolimus (tacrolimus).

[020引憐酸酶抑制剂的实例包括:BN82002盐酸盐、CP-91149、花專海绵诱癌素A (calyculin A)、斑莖酸(cantharidic acid)、斑莖素(cantharidin)、氯氯菊醋kypermethrin)、乙基-3,4-迪憐他汀(eth}d-3,4-dephostatin)、福司曲星(foshiecin) 钢盐、MAZ51、甲基-3,4-迪憐他汀(methyl-3,4-dephostatin)、NSC 95397、去甲斑莖素(norcantharidin)、来自凹形原甲藻(prorocentrum concavum)的冈田酸锭盐、冈田酸、冈田酸钟盐、冈田酸钢盐、氧化苯肿、各种憐酸酶抑制剂混合、蛋白憐酸酶1C、蛋白憐酸酶2A抑制蛋白、蛋白憐酸酶2A1、蛋白憐酸酶2A2、原饥酸钢。 [Examples 020 Rei lead acid inhibitors include: BN82002 hydrochloride, CP-91149, special flower calyculin A (calyculin A), stem spot acid (cantharidic acid), spot stem element (cantharidin), chlorobenzyl chloride Ju vinegar kypermethrin), ethyl-3,4-di pity statins (eth} d-3,4-dephostatin), Fu 18 Melpomene (foshiecin) steel salt, MAZ51, methyl-3,4-di pity statins (methyl -3,4-dephostatin), NSC 95397, norepinephrine spot stem element (norcantharidin), okadaic acid salts of the ingot from the female Prorocentrum (prorocentrum concavum) of okadaic acid, okadaic acid salts clock, steel okadaic acid salts, swelling oxidation of benzene, mixing the various pity acid inhibitor, acids pity enzyme protein 1C, protein 2A pity glucuronidase inhibiting protein, protein pity acid enzymes 2A1, 2A2 enzyme protein pity acid, acid hunger original steel.

[0209] 在一些实施例中,在此所描述的抗原也与合成纳米载体偶联。 [0209] In some embodiments, the antigen described herein can also be conjugated to synthetic carrier nm. 在一些实施例中,运些抗原与运些免疫抑制剂偶联至其上的相同或不同的合成纳米载体偶联。 In some embodiments, the antigen and transport some of these transport inhibitor to immunoconjugate thereof on the same or different conjugated synthetic nanocarriers. 在其他实施例中,运些抗原不与任何合成纳米载体偶联。 In other embodiments, these antigens are not conjugated to transport any synthetic nanocarriers. 运些抗原包括在此提供的任何抗原,或其片段或衍生物,此类抗原与炎性、自身免疫性疾病、过敏、器官或组织排斥、移植物抗宿主病有关联;移植抗原W及治疗性蛋白抗原。 These antigens include any antigen transport, or a fragment or derivative thereof provided herein, such antigens associated with inflammatory, autoimmune disease, allergy, organ or tissue rejection, graft versus host disease associated; W transplantation antigens and treatment protein antigen. 可W从所提供的或本领域中另外已知的任何抗原中获得或衍生出多个表位、或包含运些表位的蛋白质、多肤或肤。 W may be any antigen known additionally be obtained from or provided in the present art or more epitopes derived from, or comprising these epitopes transport protein, a skin or skin.

[0210] 治疗性蛋白包括但不限于:可输注的治疗性蛋白、酶、酶辅助因子、激素、凝血因子、细胞因子和干扰素、生长因子、单克隆抗体、及多克隆抗体(例如,作为替代疗法向受试者给予的多克隆抗体)、W及与庞贝氏病相关的蛋白质(例如,阿葡糖巧酶a、rhGAA(例如, Myozyme和Lumizyme(健赞公司(Genzyme)))。治疗性蛋白还包括设及在凝血级联系统中的蛋白质。治疗性蛋白包括但不限于:因子VIII、因子VII、因子IX、因子V、血管性血友病因子、 von化Idebrant因子、组织型纤溶酶原激活物、膜岛素、生长激素、α促红细胞生成素、VEGF、 促血小板生成素、溶菌酶、抗凝血酶等等。治疗性蛋白还包括脂肪因子类,如瘦素和脂联素。 如下和在此其他地方描述了治疗性蛋白的其他实例。还包括作为表位、包含该表位的蛋白质、多肤或肤而提供的任何运些治疗性蛋白的片 [0210] Therapeutic proteins include, but are not limited to: therapeutic infusible proteins, enzymes, enzyme cofactors, hormones, blood clotting factors, interferons, and cytokines, growth factors, monoclonal antibodies, and polyclonal antibodies (e.g., as a polyclonal antibody replacement therapy administered to a subject), W Pompe disease and related proteins (e.g., gluconic a clever enzymes a, rhGAA (e.g., Myozyme and Lumizyme (Genzyme (Genzyme))) therapeutic proteins and protein is provided comprising a further therapeutic protein in the blood coagulation cascade include, but are not limited to: factor VIII, factor VII, factor IX, factor V, von Willebrand factor, von of Idebrant factor, tissue type plasminogen activator, film islands, growth hormone, [alpha] erythropoietin, VEGF, thrombopoietin, lysozyme, antithrombin, etc. therapeutic proteins include fatty further factors, such as leptin and adiponectin. follows and elsewhere herein describe other examples of therapeutic proteins. these further comprises sheet transportation any therapeutic protein as an epitope, a protein comprising the epitope, or multiple skin skin provided 段或衍生物。 Segment or derivative thereof.

[0211] 在具有溶酶体胆积症的受试者的酶替代疗法中使用的治疗性蛋白的实例包括但不限于:用于治疗高雪病的伊米巧酶(例如,CEREZYME™)、用于治疗法布里病的a-半乳糖巧酶A(a-gal A)(例如,阿加糖酶i3、FABRYZYME™)、用于治疗庞贝氏病的酸性a-葡糖巧酶(GAA) (例如,阿葡糖巧酶〇、0^12¥16了》、1¥02¥16了》)、用于治疗粘多糖病的芳基硫酸醋酶8(例如, 拉罗尼酶(13的〇1(1336)、41^0抓42¥]\^了《、艾度硫酸醋酶、61^^^456了《、芳基硫酸醋酶8、 NAGLAZYME™) 〇 [0211] Examples of therapeutic proteins for use in enzyme replacement therapy in a subject having lysosomal volume bladder disorders include, but are not limited to: for the treatment of Gaucher's disease imiglucerase Qiao enzymes (e.g., CEREZYME ™), a- galactose for treating Fabry's disease Qiao enzyme a (a-gal a) (e.g., agalsidase i3, FABRYZYME ™), for the treatment of Pompe disease acid Qiao enzyme glucose a- (GAA ) (e.g., gluconate a clever enzyme billion, 0 ^ 12 ¥ 16 of ", 1 ¥ 02 ¥ 16 of"), arylsulfatase useful for the treatment of mucopolysaccharidosis esterase 8 (e.g., an enzyme Laluo Ni (13 the 〇1 (1336), 41 catch 42 is ¥ ^ 0] \ ^ a ", Aldo acid esterase, ^^^ 456 61 a", arylsulfatase esterase 8, NAGLAZYME ™) square

[0212] 酶的实例包括氧化还原酶、转移酶、水解酶、裂解酶、异构酶、W及连接酶。 [0212] Examples of enzymes include oxidoreductases, transferases, hydrolases, lyases, isomerases, W, and ligase.

[0213] 激素的实例包括:稱黑素(N-乙酷基-5-甲氧基色胺)、血清素、甲状腺素(或四舰甲状腺原氨酸)(一种甲状腺激素)、Ξ舰甲状腺原氨酸(一种甲状腺激素)、肾上腺素化pine地rine)(或肾上腺素(a化enaline))、去甲肾上腺素(Nor邱in邱brine)(或去甲肾上腺素(nora化enaline))、多己胺(或催乳素抑制激素)、抗苗勒氏管激素(或苗勒氏管抑制因子或激素)、脂联素、促肾上腺皮质激素(A化enocorticotropic hormone)(或促肾上腺皮质激素(corticotropin))、血管紧张素原和血管紧张素、抗利尿激素(Anti diuretic hormone)(或加压素(vasopressin)、精氨酸加压素(a;rginine vasopressin))、屯、房利钢肤(或屯、房肤)、降巧素、缩胆囊素(化olecystokinin)、促肾上腺皮质激素释放激素、红细胞生成素、促卵泡激素、胃泌素、胃饥饿素(Ghrelin)、膜高血糖素(Glucagon)、膜高血糖素样肤(GLP-1)、GIP、促性 Examples [0213] Hormone comprising: said melanin (N- ethyl-5-methoxytryptamine cool), serotonin, thyroxine (or four-ship triiodothyronine) (a kind of thyroid hormone), a Cascade ship thyroid triiodothyronine (s thyroid hormone), pine of epinephrine to Rine) (or epinephrine (a of enaline)), noradrenaline (Nor Qiu Qiu in brine) (or norepinephrine (Nora of enaline) ), multi-hexylamine (or prolactin inhibiting hormone), an anti-Mullerian hormone (or Mullerian inhibiting factor or hormone), adiponectin, adrenocorticotropic hormone (A of enocorticotropic hormone) (or adrenocorticotropic hormones (in Corticotropin)), angiotensinogen and angiotensin, antidiuretic hormone (anti diuretic hormone) (or vasopressin (vasopressin was), arginine vasopressin (a; rginine vasopressin)), Tun, Fannie steel skin (or village, skin room), clever drop hormone, cholecystokinin (of olecystokinin), corticotropin releasing hormone, erythropoietin, follicle stimulating hormone, gastrin, ghrelin (of ghrelin), high film glucagon (glucagon), glucagon-like film skin (GLP-1), GIP, gonadotropin 腺激素释放激素、生长激素释放激素、人绒毛膜促性腺激素、人胎盘催乳素、生长激素、抑制素(InWbin)、膜岛素、膜岛素样生长因子(或生长调节素)、瘦素、促黄体激素、促黑素细胞激素、阿立新(Orexin )、催产素、甲状旁腺激素、催乳素、松弛素、分泌素、 促生长素抑制素、促血小板生成素、促甲状腺激素(或促甲状腺素)、促甲状腺素释放激素、 皮质醇、醒固酬、睾酬、脱氨表雄酬、雄締二酬、二氨睾酬、雌二醇、雌酬、雌Ξ醇、黄体酬、骨化Ξ醇(1,25-二径基维生素D3)、骨化二醇(25-径基维生素D3)、前列腺素类、白Ξ締类、前列腺环素、凝血曝焼、催乳素释放激素、促脂解素、脑利钢肤、神经肤Y、组胺、内皮素、膜多肤、肾素、W及脑啡肤。 Thyroid hormone releasing hormone, growth hormone releasing hormone, human chorionic gonadotropin, human placental lactogen, growth hormone, inhibin (InWbin), membrane Insulin, Insulin-like growth factor film (or somatomedin), leptin , luteinizing hormone, melanocyte stimulating hormone, orexin (of orexin), oxytocin, parathyroid hormone, prolactin, relaxin, secretin, somatostatin, thrombopoietin, thyroid stimulating hormone (or TSH), thyrotropin-releasing hormone, cortisol, wake solid paid, testis paid, DHEA paid deaminase, two male association pay, pay diamine testis, estradiol, pay female Ξ alcohol, luteinizing reward , Ξ alcohol ossification (1,25-vitamin D3 diameter yl), calcifediol (25-yl diameter vitamin D3), prostaglandins, white Ξ associative classes, prostacyclin, thromboxanes exposed firing, prolactin releasing hormone, lipotropin, brain steel skin benefit, neuropeptide Y, histamine, endothelin, multiple membrane skin, renin, W, and brain endorphins.

[0214] 血液和凝血因子的实例包括:因子1(纤维蛋白原)、因子11(凝血酶原)、组织因子、 因子V(促凝血球蛋白原、易变因子)、因子VII(稳定因子、前转变素)、因子VIII(抗血友病球蛋白)、因子IX(克雷司马斯因子(Christmas factor)或血浆促凝血酶原激酶组分)、因子X (斯图亚特因子(S^art-Prower factor))、因子Xa、因子XI、因子XII(哈格曼因子巧ageman factor))、因子ΧΙΙΚ纤维蛋白稳定因子)、血管性血友病因子、前激肤释放酶(费莱彻因子(Fletcher factor))、高分子量激肤原(HMWK)(菲茨杰拉德因子)、纤连蛋白、纤维蛋白、凝血酶、抗凝血酶III、肝素辅助因子II、蛋白C、蛋白S、蛋白Z、蛋化相关蛋白酶抑制剂(ZPI)、 纤溶酶原、02-抗纤维蛋白溶酶、组织型纤溶酶原激活物(tPA)、尿激酶、纤溶酶原激活物抑审IJ剂-l(PAIl)、纤溶酶原激活物抑制剂-2(PAI2)、癌性促凝物质、W及阿 [0214] Examples of blood clotting and factors include: 1 factor (fibrinogen), factor 11 (prothrombin), tissue factor, factor V (proaccelerin, labile factor), factor VII (factor stability, proconvertin), factor VIII (antihemophilic globulin), factor IX (factor Clay Division Maas (Christmas factor) or plasma thromboplastin component), factor X (Stuart factor (S ^ art-Prower factor)), factor Xa, factor XI, factor XII (Hageman factor Qiao ageman factor)), factor ΧΙΙΚ fibrin-stabilizing factor), von Willebrand factor, stimulated skin before releasing enzyme (Feilai Che factor (Fletcher factor)), the original skin excited molecular weight (HMWK) (Fitzgerald factor), fibronectin, fibrin, thrombin, antithrombin III, heparin cofactor II, protein C, protein S , protein Z, egg correlation protease inhibitor (ZPI), plasminogen, antiplasmin 02- enzyme, tissue plasminogen activator (of tPA), urokinase, plasminogen activator inhibition Laid IJ agent -l (PAIl), plasminogen activator inhibitor -2 (PAI2), cancer procoagulant, W and A 法依伯汀(益比奥化pogen)、普罗克里特(Procrit))。 Epoetin alfa (gain of Biot pogen), general Crete (Procrit)).

[0215] 细胞因子的实例包括:淋己因子、白细胞介素、及趋化因子、1型细胞因子(如IFN- γ、TGF-目)、1^及2型细胞因子(如比-4、比-10、W及比-13)。 Examples [0215] Cytokines include: hexyl leaching factor, interleukins, and chemokines, Type 1 cytokines (e.g., IFN- γ, TGF- mesh), ^ 1 and type 2 cytokines (e.g., ratio of -4, than -10, W ratio and -13).

[0216] 生长因子的实例包括:肾上腺髓质素(AM)、血管生成素(Ang)、自分泌运动因子、骨形态发生蛋白(BMP)、脑源性神经营养因子(BDNF)、表皮生长因子化GF)、红细胞生成素化P0)、成纤维细胞生长因子(FGF)、胶质细胞系源性神经营养因子(GDNF)、粒细胞集藩刺激因子(G-CSF)、粒细胞巨日篮细胞集藩刺激因子(GM-CSF)、生长分化因子-9(GDF9)、肝细胞生长因子化GF)、肝细胞瘤源性生长因子化DGF)、膜岛素样生长因子(IGF)、促移行因子、肌生长抑制素(GDF-8)、神经生长因子(NGF)和其他神经营养蛋白、血小板源性生长因子(PDGF)、 促血小板生成素(ΤΡ0)、α-转化生长因子(TGF-a)、目-转化生长因子(TGF-目)、α-肿瘤坏死因子(TNF-a)、血管内皮生长因子(VEGF)、Wnt信号通路、胎盘生长因子(P1GF)、[(胎牛生长激素)](FBS)、比-1、比-2、比-3、比-4、比-5、比-6、^及比-7。 [0216] Examples of growth factors include: adrenomedullin (AM), angiopoietin (of Ang), autocrine motility factor, bone morphogenetic protein (BMP), brain-derived neurotrophic factor (of BDNF), epidermal growth factor of GF), erythropoietin of P0), fibroblast growth factor (FGF), glial cell line-derived neurotrophic factor (GDNF), granulocyte-stimulating factor fan (G-CSF), granulocyte Juri basket Phan cell assemblies stimulating factor (GM-CSF), growth differentiation factor -9 (GDF9), hepatocyte growth factor of GF), hepatoma derived growth factor of DGF), membrane Insulin-like growth factor (IGF), pro migration factor, myostatin (GDF-8), nerve growth factor (NGF) and other neurotrophins, platelet derived growth factor (PDGF), thrombopoietin (ΤΡ0), α- transforming growth factor (TGF- [beta A), mesh - transforming growth factor (TGF- [beta mesh), tumor necrosis factor alpha] (TNF-a), vascular endothelial growth factor (VEGF), Wnt signaling pathway, placental growth factor (P1GF), [(fetal bovine somatotropin )] (FBS), than -1, -2 ratio than -3, -4 ratio than -5, -6 ratio, and the ratio ^ -7.

[0217] 单克隆抗体的实例包括:阿己伏单抗(Abagovomab)、阿昔单抗(Abciximab)、阿达木单抗(Adalimumab)、阿德木单抗(Adecatumumab)、阿非莫单抗(Afelimomab)、阿夫上珠(Afutuzumab)、培化阿珠单抗(Alacizumab pegol)、ALD、阿仑单抗(Alemtuzumab)、喷替酸阿妥莫单抗(Altumomab pentetate)、麻安莫单抗(Anatumomab mafenatox)、安芦珠单抗(Anrukinzumab)、抗胸腺细胞球蛋白、阿泊珠单抗(Apol izumab)、阿西莫单抗(Arcitumomab)、阿塞珠单抗(Aselizumab)、那他珠单抗(Atlizumab)(托珠单抗(tocilizumab))、阿托木单抗(Atorolimumab)、己品珠单抗(Bapineuzumab)、己利昔单抗(Basiliximab)、己维昔单抗(Bavituximab)、贝妥莫单抗(Bectumomab)、贝利木单抗(Belimumab)、贝那利珠单抗(Benralizumab)、柏替木单抗(Beftilimumab)、贝索单抗(Besilesomab)、贝伐单抗(Bevacizumab)、比西单抗(Biciromab)、比伐珠单抗-美登素(Bivatuzumabmertansine) [0217] Examples of monoclonal antibodies include: A monoclonal antibody has V (Abagovomab), abciximab (Abciximab), adalimumab (Adalimumab), Adelaide adalimumab (Adecatumumab), afelimomab ( Afelimomab), d'beads (Afutuzumab), culture of Azhu mAb (alacizumab pegol), ALD, alemtuzumab (alemtuzumab), pentetic acid a properly Mo mAb (Altumomab pentetate), hemp Anmo mAb (Anatumomab mafenatox), Lu Ann trastuzumab (Anrukinzumab), anti-thymocyte globulin, A Park natalizumab (Apol izumab), ASIMO monoclonal antibody (Arcitumomab), Azeri natalizumab (Aselizumab), he natalizumab (atlizumab) (tocilizumab (tocilizumab)), Alto adalimumab (Atorolimumab), has products natalizumab (Bapineuzumab), has infliximab (Basiliximab), has dimension infliximab (Bavituximab ), Tony Mok properly monoclonal antibody (Bectumomab), belimumab (Belimumab), Tony Canary natalizumab (Benralizumab), cypress wood for monoclonal antibody (Beftilimumab), Besso monoclonal antibody (Besilesomab), bevacizumab anti- (Bevacizumab), than anti Xidan (Biciromab), than Avastin - maytansine (Bivatuzumabmertansine) 兰妥莫单抗(Blinatumomab)、贝伦妥单抗-维多汀(Bren化ximab vedotin)、布雷奴单抗(Briakinumab)、卡那单抗(Canakinumab)、莫坎妥珠单抗(Cantuzumab me:rtansine)、卡罗单抗喷地版(Capromab pendetide)、卡妥索单抗Watumaxomab)、西利珠单抗(Cedelizumab)、赛妥珠单抗(Certolizumab pegol)、西妥昔单抗(Cetuximab)、泊西他珠单抗(Citatuzumab bogatox)、西妥木单抗(Cixutumumab)、克立昔单抗(Clenoliximab)、克伐珠单抗(Clivatuzumabtetraxetan)、可那木单抗(Conatumumab)、达西珠单抗(Dacetuzumab)、达利珠单抗(Daclizumab)、达雷木单抗(Daratumumab)、地诺单抗(Denosumab)、地莫单抗(Detumomab)、阿托度单抗(Dorlimomab aritox)、达利珠单抗(Dorlixizumab)、依美昔单抗(Ecromeximab)、依库珠单抗(Eculizumab)、埃己单抗化dobacomab)、依决洛单抗化drecolomab)、依法利珠单抗(Efalizumab)、依芬古单抗(Efungumab)、依洛珠单抗(Elo tuzumab Lan Mo properly monoclonal antibody (Blinatumomab), Belem trastuzumab - Wei Duoting (Bren of ximab vedotin), Bray slave monoclonal antibody (Briakinumab), kanamycin monoclonal antibody (Canakinumab), Mokan trastuzumab (Cantuzumab me : rtansine), Carlo monoclonal antibody to spray version (capromab pendetide), Catumaxomab monoclonal antibody Watumaxomab), Sealy natalizumab (Cedelizumab), certolizumab (certolizumab pegol), cetuximab (cetuximab) , Park West natalizumab (Citatuzumab bogatox), cetuximab adalimumab (cixutumumab), Kerry infliximab (Clenoliximab), g bevacizumab (Clivatuzumabtetraxetan), but that adalimumab (conatumumab), Darcy natalizumab (Dacetuzumab), daclizumab (daclizumab), Darrell adalimumab (Daratumumab), denosumab (Denosumab), the Mo monoclonal antibody (Detumomab), Alto degree monoclonal antibody (Dorlimomab aritox) , daclizumab (Dorlixizumab), according to the US infliximab (Ecromeximab), eculizumab (eculizumab), Egypt had mAb of dobacomab), edrecolomab of drecolomab), efalizumab (Efalizumab), Eve ancient monoclonal antibody (Efungumab), according to Luo natalizumab (Elo tuzumab )、艾西莫单抗(Elsilimomab)、培化恩莫单抗(Enlimomab pegol )、西艾匹莫单抗(Ep it umomab cituxetan)、依帕珠单抗(Epra tuzumab )、厄利珠单抗(Erl izumab)、厄妥索单抗(Ertumaxomab)、埃达珠单抗化taracizumab)、艾维单抗化xbivirumab)、法索单抗(Fanolesomab)、法控莫单抗(Faralimomab)、法利珠单抗(Farletuzumab)、泛维珠单抗(Felvizumab)、非扎奴单抗(Fezakinumab)、芬妥木单抗(Figitumumab)、芳妥珠单抗(Fontolizumab)、福控韦单抗(Foravirumab)、夫苏木单抗(Fresolimumab)、加利昔单抗(Galiximab)、罗氏单抗(Gantenerumab)、加维莫单抗(Gavilimomab)、吉妥珠单抗-奥P坐米星(Gemtuzumabozogamicin)、GC1008、吉瑞昔单抗(Giren化ximab)、格雷帕珠单抗-维德汀(Glembatumumab vedotin)、戈利木单抗(Golimumab)、戈利昔单抗(Gomiliximab)、伊己珠单抗Qbalizumab)、替伊莫单抗Qbritumomab tiuxetan)、伊戈伏单抗(Igovomab)、英西单抗(Imciromab)、 ), Yixi Mo monoclonal antibody (Elsilimomab), training of Enmo monoclonal antibody (Enlimomab pegol), Sierra Atlantic pimozide monoclonal antibody (Ep it umomab cituxetan), epratuzumab (Epra tuzumab), Early natalizumab (Erl izumab), Eritrea proper cable mAb (Ertumaxomab), daclizumab Ada of taracizumab), Ivy mAb of xbivirumab), Faso mAb (Fanolesomab), Mo process control mAb (Faralimomab), Farley natalizumab (Farletuzumab), pan-dimensional natalizumab (Felvizumab), non-tie slave monoclonal antibody (Fezakinumab), Finland properly adalimumab (Figitumumab), aryl trastuzumab (fontolizumab), Fu Wei control mAb (Foravirumab ), Fu Su adalimumab (Fresolimumab), Gary infliximab (Galiximab), Roche monoclonal antibody (Gantenerumab), Jiawei Mo monoclonal antibody (Gavilimomab), gemtuzumab - Austria P rice sit Star (Gemtuzumabozogamicin) , GC1008, Jerry infliximab (Giren of ximab), Ge Leipa natalizumab - Weide Ting (Glembatumumab vedotin), golimumab (golimumab), golimumab infliximab (Gomiliximab), Iraq has a single bead anti Qbalizumab), ibritumomab tiuxetan Qbritumomab tiuxetan), Andre V monoclonal antibody (Igovomab), British anti Xidan (Imciromab), 英夫利昔单抗(Inf liximab)、英妥木单抗(Intetumumab)、伊诺莫单抗(Inolimomab)、伊珠单抗-奥加米星(Ino化zumab ozogamicin)、伊匹单抗(Ipilimumab)、伊妥木单抗(Iratumumab)、凯利昔单抗化eliximab)、控贝珠单抗(Labetuzumab)、来金珠单抗化ebrikizumab)、来马索单抗化emalesomab)、乐德木单抗化erdelimumab)、来沙木单抗(Xexatumumab)、利韦单抗(Xibivirumab)、林妥珠单抗(Xintuzumab)、罗佛珠单抗-美登素(Lorvotuzumab mertansine)、鲁卡木单抗(Lucatumumab)、鲁昔单抗(Lumiliximab)、马中白木单抗(Mapatumumab)、马司莫单抗(Maslimomab)、马妥珠单抗(Matuzumab)、美泊利单抗(Mepolizumab)、美替木单抗(Metelimumab)、米控珠单抗(Milatuzumab)、明瑞莫单抗(Minretumomab)、米妥莫单抗(Mitumomab)、莫罗木单抗(Morolimumab)、莫维珠单抗(Motavizumab)、莫罗单抗-CD3(Muromonab-CD3)、他那可洛单抗(Nacolomab tafenatox)、 他那莫单抗(Naptum Infliximab (Inf liximab), English properly adalimumab (Intetumumab), Yi Nuomo monoclonal antibody (Inolimomab), daclizumab Iraq - Ao Jiami star (Ino of zumab ozogamicin), ipilimumab (Ipilimumab ), Iraq proper adalimumab (Iratumumab), Kelly infliximab of eliximab), Tony control natalizumab (labetuzumab), lebrikizumab of ebrikizumab), to Marceau monoclonal antibody of emalesomab), Germany wooden music single inhibitors erdelimumab), lexatumumab (Xexatumumab), Liberi monoclonal antibody (Xibivirumab), lintuzumab (Xintuzumab), Luo beads monoclonal antibody - maytansinoid (Lorvotuzumab mertansine), Lucas adalimumab ( Lucatumumab), lumiliximab (lumiliximab), Malaysia Shiraki monoclonal antibody (Mapatumumab), Ma Secretary Mo monoclonal antibody (Maslimomab), horse trastuzumab (matuzumab), mepolizumab (mepolizumab), the United States for wood monoclonal antibody (Metelimumab), meter control natalizumab (milatuzumab), Ming Ruimo monoclonal antibody (Minretumomab), mitumomab (mitumomab), Morrow adalimumab (Morolimumab), motavizumab (Motavizumab) Morrow monoclonal antibody -CD3 (muromonab-CD3), his Crow monoclonal antibody (Nacolomab tafenatox), his MO monoclonal antibody (Naptum omabΘstafenatox)、那他珠单抗(Natalizumab)、奈己库单抗(Nebacumab)、奈昔木单抗(Necitumumab)、奈瑞莫单抗(Nerelimomab)、尼妥珠单抗(Nimotuzumab)、莫诺非莫单抗(Nofetumomab merpentan)、奥ί請王朱单抗(Ocrelizumab)、奥度莫单抗(Odulimomab)、奥法木单抗(Ofatumumab)、奥控雷单抗(Olaratumab)、奥马珠单抗(Omalizumab)、莫奥珠单抗(Opoi'tuzumab monatox)、奥戈伏单抗(Oregovomab)、奥昔珠单抗(Otelixizumab)、帕吉昔单抗(Pagibaximab)、帕利珠单抗(Palivizumab)、帕尼单抗(Panitumumab)、帕诺库单抗(Panobacumab)、帕考珠单抗(Pascolizumab)、帕尼单抗(Pemtumomab)、帕妥珠单抗(Pertuzumab)、培克珠单抗(Pexelizumab)、平妥单抗(Pintumomab )、普立昔单抗(Priliximab)、普托木单抗(Pr i tumumab )、瑞非韦鲁(Raf ivirumab)、礼来单抗(Ramucirumab)、来尼珠单抗(Ranibizumab)、瑞希己库(Raxibacumab)、瑞加韦单抗(Regavirumab)、瑞利珠单 omabΘstafenatox), natalizumab (Natalizumab), Chennai has library monoclonal antibody (Nebacumab), necitumumab (Necitumumab), Nai Ruimo monoclonal antibody (Nerelimomab), nimotuzumab (Nimotuzumab), non-Monod Mo monoclonal antibody (Nofetumomab merpentan), Austria ί king Zhu monoclonal antibody (Ocrelizumab), Austria degrees Mo monoclonal antibody (Odulimomab), ofatumumab (ofatumumab), Austrian-controlled mine monoclonal antibody (Olaratumab), omalizumab (omalizumab ), Moao natalizumab (Opoi'tuzumab monatox), oregovomab (oregovomab), oxybutynin natalizumab (otelixizumab), pagibaximab (pagibaximab), palivizumab (palivizumab), panitumumab (panitumumab), Pompano library monoclonal antibody (Panobacumab), Pascolizumab (Pascolizumab), panitumumab (pemtumomab), pertuzumab (pertuzumab), pexelizumab (pexelizumab ), flat trastuzumab (Pintumomab), PolyOne infliximab (Priliximab), Maputo adalimumab (Pr i tumumab), Swiss non-Wei Lu (Raf ivirumab), Lilly mAb (ramucirumab), to Nyidrol monoclonal antibody (Ranibizumab), Mizuki has library (raxibacumab), Rui Jiawei monoclonal antibody (Regavirumab), Rayleigh single bead 抗(Reslizumab)、利妥木单抗(Rilotumumab)、利妥珠单抗(Rituximab)、罗妥木单抗(Robatumumab)、罗利珠单抗(Rontalizumab)、罗韦来单抗(Rovelizumab)、芦利珠单抗(Ruplizumab)、喷地版沙妥莫单抗(Satumomab pendetide)、司韦单抗(Sevirumab)、西罗珠单抗(SiLbro化zumab)、西法木单抗(Sifalimumab)、司妥昔单抗(Siltuximab)、希普利珠单抗(Siplizumab)、苏兰珠单抗(Solanezumab)、索尼普单抗(Sonepcizumab)、索上珠单抗(Sontuzumab)、司他芦单抗(Stamulumab)、硫索单抗(Sulesomab)、他珠单抗(Tacatuzumab tetraxetan)、他度珠单抗(Tadocizumab)、他利珠单抗(Talizumab)、他尼珠单抗(Tanezumab)、帕他莫单抗(T'aplitumomab paptox)、特非珠单抗(Teflbazumab)、阿替莫单抗(Telimomab aritox)、替妥莫单抗(Tenatumomab)、替奈昔单抗(Teeneliximab)、替利珠单抗(Teplizumab)、替西莫单抗(Ticilimumab)(替西木单抗(tremelimumab))、替加珠单 Anti- (Reslizumab), rilotumumab (Rilotumumab), rituximab, trastuzumab (Rituximab), Luo proper adalimumab (Robatumumab), Raleigh natalizumab (Rontalizumab), Grove to monoclonal antibody (Rovelizumab), Lu Li Zhu monoclonal antibody (Ruplizumab), spray the sand proper version Mo monoclonal antibody (Satumomab pendetide), Secretary Wei monoclonal antibody (Sevirumab), Ciro natalizumab (SiLbro of zumab), sifalimumab (sifalimumab), Division of rituximab alone anti (Siltuximab), Shipley daclizumab (Siplizumab), Su Lan daclizumab (solanezumab), Suoni Pu mAb (Sonepcizumab), daclizumab ropes (Sontuzumab), orlistat Lo mAb (Stamulumab), sulfur cable monoclonal antibody (Sulesomab), natalizumab (Tacatuzumab tetraxetan), he of natalizumab (Tadocizumab), his interest natalizumab (Talizumab), Thani natalizumab (Tanezumab), Pa tamoxifen monoclonal antibody (T ' aplitumomab paptox), special non-trastuzumab (Teflbazumab), temozolomide A monoclonal antibody (Telimomab aritox), for proper Mo monoclonal antibody (Tenatumomab), Chennai for infliximab (Teeneliximab), teplizumab (Teplizumab), Seamus for monoclonal antibody (ticilimumab) (for West adalimumab (tremelimumab)), for a single bead plus (Tigatuzumab)、托珠单抗(Tocilizumab)(阿替珠单抗(atlizumab))、托利珠单抗(Toralizumab)、托西莫单抗(Tositumomab)、曲妥珠单抗(Trastuzumab)、曲美木单抗(Tremelimumab)、西莫白细胞介素单抗(Tucotuzumab celmoleukin)、妥韦单抗(Tuvirumab)、乌珠单抗(Urtoxazumab)、优特克单抗(Ustekinumab)、伐利昔单抗(Vapaliximab)、维多珠单抗(Vedolizumab)、维妥珠单抗(Veltuzumab)、维帕莫单抗(Vepalimomab)、维西珠单抗(Visilizumab)、伏洛昔单抗(Volociximab)、伏妥莫单抗(Votumumab)、扎鲁木单抗(Zalutumumab)、扎木单抗(Zanolimumab)、齐拉木单抗(Ziralimumab)、W及阿佐莫单抗(Zolimomab aritox)。 (Tigatuzumab), tocilizumab (Tocilizumab) (atenolol trastuzumab (atlizumab)), Toledo natalizumab (Toralizumab), tositumomab (Tositumomab), Herceptin (Trastuzumab), song US adalimumab (Tremelimumab), Seamus interleukin monoclonal antibody (Tucotuzumab celmoleukin), duly Wei monoclonal antibody (Tuvirumab), Ukraine natalizumab (Urtoxazumab), ustekinumab (ustekinumab), cutting infliximab (Vapaliximab), vedolizumab (vedolizumab), veltuzumab (Veltuzumab), Weipa Mo monoclonal antibody (Vepalimomab), Weixi natalizumab (Visilizumab), V Los infliximab (volociximab), V Mo properly monoclonal antibody (Votumumab), zalutumumab (zalutumumab), zanolimumab (zanolimumab), Zillah adalimumab (Ziralimumab), W and A Zuomo monoclonal antibody (Zolimomab aritox).

[0218] 输注疗法或可注射的治疗性蛋白的实例包括:例如,塔西单抗(Tocilizumab)(罗氏公司(RocheVActemra發)、日-1抗膜蛋白酶(卡美达公司化amadaVAAT)、Hema削e敏(Affymax和Takeda,合成肤)、白蛋白干扰素a-2b (诺华公司(NovartisVZalbinT™)、 Rhucin® (嫁接集团(Pharming Group),Cl抑制剂替代疗法)、替莫瑞林(tesamorelin)(治疗技术公司(TheratechnologiesVEgrifta,合成生长激素释放因子)、奥瑞珠单抗(ocrelizumab)(遗传技术公司(Genentech)、罗氏公司(Roche)W及生物遗传公司(Biogen))、杯利单抗(belimumab)(葛兰素史克公司(GlaxoSmithKline), Benlysta般)、聚乙二醇重组尿酸酶(Savient制药公司/KirstexxaT™)、α-他利巧酶(?1'〇1日1;[义公司/ 化1730)、阿加糖酶0(夏尔公司(化^6)/化01)13龄1(巧)、葡糖脑巧脂酶0(夏尔公司(511山6))。 Examples of infusion therapy or injectable therapeutic protein [0218] includes: for example, monoclonal antibody Tacitus (Tocilizumab) (Roche (RocheVActemra fat), anti-days-1 membrane protease (Mita card of amadaVAAT), Hema cut e Min (Affymax and Takeda, synthetic skin), albumin, interferon a-2b (Novartis (NovartisVZalbinT ™), Rhucin® (grafting Group (Pharming Group), Cl inhibitor replacement therapy), for Morui Lin (tesamorelin) (treatment technologies, Inc. (TheratechnologiesVEgrifta, synthetic growth hormone releasing factor), ocrelizumab (ocrelizumab) (genetic technologies, Inc. (Genentech), Roche (Roche) W and Biogen (Biogen)), Lee cup monoclonal antibody ( belimumab) (GSK (GlaxoSmithKline), Benlysta like), polyethylene glycol recombinant uricase (Savient Pharmaceuticals / KirstexxaT ™), α- he skillfully Lee enzyme (1 '1, 1 billion;? [Yi company / of 1730), agalsidase 0 (Charles company (of ^ 6) / of 01) 13 age 1 (clever), glucocerebrosidase clever lipase 0 (Charles company (511 mountain 6)).

[0219] 对根据本发明的方面有用的另外的治疗性蛋白对于本领域普通技术人员而言将是显而易见的,并且本发明在此方面是不受限的。 [0219] In addition to therapeutic proteins useful in accordance with aspects of the present invention to those of ordinary skill in the art will be apparent, and the invention is not limited in this regard.

[0220] 在一些实施例中,可W对一种组分(如抗原或免疫抑制剂)进行分离。 [0220] In some embodiments, it may be one component of W (e.g., an antigen or an immunosuppressant) isolated. 分离是指该成分与它的天然环境分离并且W容许对它进行鉴定或使用的足够量存在。 It means that the isolated component is separated from its native environment and W allowable amount sufficient identification or use it is present. 运意味着,例如, 该成分可WQ)通过表达克隆选择性地产生或(ii)如通过色谱法或电泳进行纯化。 Transport means, e.g., the ingredient may be WQ) is selectively produced by expression cloning or (ii) purified as by chromatography or electrophoresis. 分离的成分可W是基本上纯的,但是不需要如此。 W is separated components may be substantially pure, but need not be so. 由于在药物制剂中分离的成分可W与药学上可接受的赋形剂相混合,按该制剂的重量计该成分可W仅包含小的百分比。 Due to the separation of the components may be W pharmaceutical formulation with a pharmaceutically acceptable excipient are mixed, the formulation by weight of the component count may comprise only a small percentage W. 虽然如此,该成分是分离的原因在于,它已经从在活系统中可能与其结合的物质中分离出来,即从其他脂质或蛋白质中分离出来。 Nevertheless, this component is separated from the reason that it has been separated from the material in a living system in connection with a possible out, i.e. isolated from other proteins or lipids. 在此提供的任何成分可分离的形式包含在运些组合物中。 Any composition provided herein comprises separable form in the operation of these compositions.

[0221 ] D.制造和使用本发明的组合物的方法和相关方法 [0221] D. Methods of making and using the compositions of the invention and related methods

[0222] 可W使用本领域中已知的各种各样的方法制备合成纳米载体。 [0222] W can be prepared using synthetic nanocarriers known in the art various methods. 例如,可W通过如纳米沉淀、使用流体通道的流动聚焦、喷雾干燥、单和双乳液溶剂蒸发、溶剂萃取、相分离、 研磨、微乳液步骤、微制造、纳米制造、牺牲层、简单和复杂凝聚法的方法、W及本领域的那些普通技术人员熟知的其他方法形成合成纳米载体。 For example, W by nano-precipitation, flow focusing fluid passage, spray drying, single and double emulsion solvent evaporation, solvent extraction, phase separation, grinding, microemulsions step, microfabrication, nanofabrication, the sacrificial layer, simple and complex method of coacervation, W, and other methods in the art are well known to those of ordinary skill in forming synthetic nanocarriers. 可替代地或另外地,已经说明了用于单分散半导体,传导性的、磁性的、有机的、W及其他纳米材料的水性和有机溶剂合成(Pellegrino (佩莱格里诺)等人,2005,小物质,1:48;Murray(莫雷)等人,2000,材料科学年度评论,30:545; W及化indade(特林达德)等人,2001,材料化学,13:3843)。 Alternatively or in addition, has been described for monodisperse semiconductor, conductive, magnetic, organic, aqueous W and other nanomaterials synthesis and an organic solvent (Pellegrino (Pellegrino) et al., 2005 little substance, 1: 48; Murray (Murray) et al., 2000, annual review of materials Science, 30: 545; W and of indade (Trindade) et al., 2001, materials Chemistry, 13: 3843). 在文献中已经描述了另外的方法(参见,例如,多布罗夫(Doubrow)编著,"医药中的微胶囊和纳米颗粒(Microcapsules and Nanoparticles in Medicine and Pharmacy)'',CRC出片反社,博卡拉顿(Boca Raton),1992;马西威兹(Mathiowitz)等人,1987,控释期刊(J. Control.Release),5:13;马西威兹(Mathiowitz)等人,1987,反应性聚合物(Reactive Polymers ),6:275;W及马西威兹(Mathiowitz)等人,1988,应用聚合物科学期刊(J.Appl .Polymer Sc i.),35:755;美国专利5578325 和6007845;?·保利赛利(P.Paolicelli)等人,"可W有效结合并递送病毒样颗粒的表面修饰的基于化GA的纳米颗粒(Surface-modified PLGA-based Nanoparticles that can Efficiently Associate and Deliver Virus-like 化rticles)",纳米医学(Nanomedicine) ,5(6):843-853 (2010))0 It has been described in the literature Additional methods (see, e.g., Duobuluofu (Doubrow) ed., "Pharmaceutical microcapsules and nanoparticles (Microcapsules and Nanoparticles in Medicine and Pharmacy) '', CRC antisocial the sheet, Boca Raton (Boca Raton), 1992; Marcy's Dry (Mathiowitz) et al., 1987, Journal of Controlled release (J. Control.Release), 5: 13; Marcy's Dry (Mathiowitz) et al., 1987, reaction polymer (Reactive polymers), 6: 275; W and Marcy Dry (Mathiowitz) et al., 1988, Journal of applied polymer Science (J.Appl .Polymer Sc i.), 35: 755; and U.S. Patent No. 5,578,325 6,007,845;? Paul Li Saili (P.Paolicelli) et al., "W may bind and deliver effective virus-like particles surface-modified nanoparticle based of GA's (surface-modified PLGA-based nanoparticles that can efficiently Associate and deliver virus -like of rticles) ", nanomedicine (nanomedicine), 5 (6): 843-853 (2010)) 0

[0223] 可W使用各种方法将各种材料封装在所令人希望的合成纳米载体中,运些方法包括但不限于:c ·阿斯泰特(C.Astete)等人,"PLGA纳米颗粒的合成和表征(Synthesis and characterization of PLGA nanoparticles)"生物材料科学期刊(J.Biomater.Sci.)聚合物版,第17卷,第3期,第247-289页(2006);K ·阿芙古斯塔基斯化.AvgoustakisrPEG化的聚(丙交醋)和聚(丙交醋-共-乙交醋)纳米颗粒:制备、特性W及在药物递送中的可能应用(Pegylated Poly(Lactide)and Poly(Lactide-Co-Glyco1ide)Nanoparticles: Preparation,Properties and Possible Applications in Drug Delivery)''当前药物递送(Current Drug DeliveryH :321-333(2004);里斯· C(C.Reis)等人,。纳米封装I.用于制备载药聚合物纳米颗粒的方法(Nanoencapsulation I.Methods for preparation of 化ug-loaded polymeric nanoparticles)"纳米医学(Nanomedicine)2:8-21(2006) ;P ·保利赛利(P.Paolicelli)等人,"可W有效结合和递 [0223] W can be a variety of materials using various methods encapsulated in the desirable synthetic nanocarriers, transported such methods include but are not limited to: c · Astete (C.Astete) et al., "PLGA nano synthesis and characterization of particles (synthesis and characterization of PLGA nanoparticles) "biological materials science journals (. J.Biomater.Sci) polymer Edition, Vol. 17, No. 3, pp. 247-289 (2006); K · a芙古斯塔基斯 .AvgoustakisrPEG of poly (lactide vinegar) and poly (lactide acetate - co - glycolide vinegar) nanoparticles: preparation, properties and potential applications in W in drug delivery (Pegylated poly (lactide) and Poly (Lactide-Co-Glyco1ide) Nanoparticles: Preparation, Properties and Possible Applications in Drug delivery) '' The current drug delivery (current Drug DeliveryH: 321-333 (2004); Rees · C (C.Reis), et al. method (nanoencapsulation I.Methods for preparation of oriented ug-loaded polymeric nanoparticles) nanoencapsulation I. for the preparation of drug-loaded polymeric nanoparticles "nanomedicine (nanomedicine) 2: 8-21 (2006); P · Li Saili Bao (P.Paolicelli) et al., "but effective combination of W and delivery 送病毒样颗粒的表面修饰的基于化GA的纳米颗粒(Surface-modified PLGA-based Nanoparticles that can Efficiently Associate and Deliver Virus-like Particles)"纳米医学(胞11〇1110(11〇1]10),5(6):843- 853(2010)。可W使用其他适合于将材料封装至合成纳米载体中的方法,运些方法包括但不限于披露在昂格尔化nger)的美国专利6,632,671 (2003年10月14日)中的方法。 Sending virus-like particle surface modified nanoparticles of the GA-based (Surface-modified PLGA-based Nanoparticles that can Efficiently Associate and Deliver Virus-like Particles) "Nanomedicine (extracellular 11〇1110 (11〇1] 10), 5 (6): 843-853 (2010) can be adapted to other W encapsulating material to the synthesized nanocarriers, such methods include but are not transported to the disclosure of U.S. Patent Unger nger) of 6,632, methods 671 (October 14, 2003) in.

[0224] 在某些实施例中,通过纳米沉淀工艺或喷雾干燥来制备合成纳米载体。 [0224] In certain embodiments, drying is prepared by synthetic nanocarriers nano-precipitation process or spray. 可W改变在制备合成纳米载体中使用的条件W产生具有所希望的大小和特性(例如,疏水性、亲水性、外部形态学、"粘性"、形状等等)的颗粒。 Condition W may be used in the preparation of synthetic change nanocarriers W generates a desired size and characteristics (e.g., hydrophobicity, hydrophilicity, external morphology, "stickiness", shape, etc.) of the particles. 制备运些合成纳米载体的方法和使用的条件(例如,溶剂、溫度、浓度、空气流速等等)可W取决于有待偶联至运些合成纳米载体上的材料和/或该聚合物基质的组成。 The method of operation and some synthetic nanocarriers conditions (e.g., solvent, temperature, concentration, air flow rate, etc.) used in the preparation of W to be coupled to the transport depends material and / or on the polymer matrix of some synthetic nanocarriers composition.

[0225] 如果通过任何W上方法制备的颗粒具有在希望的范围外的大小范围,那么可W按大小分类运些颗粒,例如使用一个筛。 [0225] If you have a size range outside of the desired range of particles prepared by the method of any of W, then W can be transported by size of these particles, for example using a sieve.

[0226] 本发明的合成纳米载体的成分(即,组分)(如构成一个免疫特征表面的部分、祀向部分、聚合物基质、表位、或含有表位的蛋白质、多肤或肤、免疫抑制剂等)可W例如通过一个或多个共价键偶联至整体的合成纳米载体上,或可W借助一个或多个连接物来偶联。 [0226] Synthesis ingredient nanocarriers of the present invention (i.e., component) (e.g., constituting a portion of the surface of immune feature, part of the Si, the polymer matrix, an epitope, or epitope-containing protein, skin or multiple skin, immunosuppressive agents, etc.) may be, for example, by one or more W covalently coupled to synthetic nanocarriers as a whole, or W via one or more linkers to couple. 使合成纳米载体功能化的另外方法可W由W下文献改编而来:萨尔茨曼(Saltzman)等人的公布的美国专利申请2006/0002852、戴斯蒙(DeSimone)等人的公布的美国专利申请2009/ 0028910、或默西(Mudhy)等人的公布的国际专利申请W0/2008/127532 A1。 Synthetic nanocarriers further functionalized method adapted from the literature W W: U.S. Pat Saltzman (Saltzman) et al., Published application 2006/0002852, Desmond (DeSimone,) et al., Published U.S. Patent application 2009 / 0,028,910, or Mersey (Mudhy) and others published international patent application W0 / 2008/127532 A1.

[0227] 可替代地或另外地,可W经由非共价相互作用使合成纳米载体直接地或间接地偶联至组分上。 [0227] Alternatively or additionally, the synthetic nanocarrier W interaction via a non-covalently conjugated directly or indirectly to the component. 在非共价的实施例中,通过非共价相互作用介导非共价偶联,运些非共价相互作用包括但不限于:电荷相互作用、亲和相互作用、金属配位、物理吸附、主-客体相互作用、 疏水相互作用、TT堆积相互作用、氨键相互作用、范德华相互作用、磁相互作用、静电相互作用、偶极-偶极相互作用、和/或其组合。 In an embodiment of the non-covalent, by non-covalent interactions mediated by non-covalent coupling, non-covalent interactions of these operation include, but are not limited to: charge interactions, affinity interactions, metal coordination, physical adsorption , host - guest interactions, hydrophobic interactions, TT stacking interactions, ammonia bonding interactions, van der Waals interactions, magnetic interactions, electrostatic interactions, dipole - dipole interactions, and / or combinations thereof. 此类偶联可W安排在本发明合成纳米载体的一个外表面或一个内表面上。 Such coupling may be arranged on a W outer surface of the synthetic nanocarrier of the present invention or an inner surface. 在多个实施例中,封装和/或吸附是偶联的形式。 In various embodiments, the package and / or adsorption is conjugated form. 在多个实施例中,本发明的运些合成纳米载体可W通过在相同媒介物或递送系统中混合而与抗原相组合。 In various embodiments, some synthetic nanocarriers operation of the present invention may be combined with W by mixing an antigen in the same vehicle or delivery system.

[0228] 可W使用传统的药物混合方法组合多个群体的合成纳米载体W形成根据本发明的药物剂型。 [0228] W may be mixed using conventional pharmaceutical method of combining a plurality of groups of synthetic nanocarriers W pharmaceutical dosage form according to the invention. 运些方法包括液体-液体混合,其中两种或更多种各自含有一个或多个纳米载体子集的悬浮液被直接组合或经由一个或多个含有稀释剂的容器被集合在一起。 Such methods include liquid transport - liquid mixing, wherein the two or more each containing one or more subsets nanocarriers suspension is combined directly or via one or more containers containing diluent are grouped together. 因为合成纳米载体还可粉末形式生产或胆存,所W可W进行干燥的粉末-粉末混合,因为可W将两种或更多种粉末再悬浮于共同的介质中。 Because the synthetic nanocarriers in powder form may also be produced or stored bile, the W powder can be dried W - powder were mixed, because the W can be two or more powder was resuspended in a common medium. 取决于纳米载体的特性和它们的相互作用潜力,一种或另一种混合途径可能也有优势。 Depending on the characteristics of nanocarriers and their potential interactions, one or another hybrid approach may have the advantage.

[0229] 包含合成纳米载体的典型的发明组合物可W包含无机或有机缓冲剂(例如,憐酸、 碳酸、乙酸、或巧樣酸的钢盐或钟盐)和抑调节剂(例如,盐酸、氨氧化钢或氨氧化钟、巧樣酸或乙酸的盐、氨基酸和它们的盐)、抗氧化剂(例如,抗坏血酸、α-生育酪)、表面活性剂(例如,聚山梨醋20、聚山梨醋80、聚氧乙締9-10壬基酪、去氧胆酸钢)、溶液和/或低溫/冻干稳定剂(例如,薦糖、乳糖、甘露醇、海藻糖)、渗透调节剂(例如,盐类或糖类)、抗细菌剂(例如, 苯甲酸、苯酪、庆大霉素)、消泡剂(例如,聚二甲基娃酬(polydimethylsilozone))、防腐剂(例如,硫柳隶、2-苯氧乙醇、EDTA)、聚合物稳定剂和粘度调节剂(例如,聚乙締化咯酬、泊洛沙姆488、簇甲基纤维素)、W及共溶剂(例如,甘油、聚乙二醇、乙醇)。 [0229] Typical compositions of the invention may comprise synthetic nanocarriers W comprises an inorganic or organic buffers (e.g., pity acid, carbonic acid, acetic acid, or an acid-like steel clever salts or salts clock) and suppressor modifiers (e.g., hydrochloric acid ammoxidation steel or ammoxidation clock, Qiao like or acetic acid salts, amino acids and their salts), antioxidants (e.g., ascorbic acid, alpha] fertility casein), a surfactant (e.g., polysorbate vinegar 20, polysorbate vinegar 80, polyoxyethylene 9-10 nonyl associative casein, steel deoxycholic acid), solutions, and / or low temperature / lyophilization stabilizer (e.g., recommended, lactose, mannitol, trehalose), osmotic adjusting agents ( For example, salts or carbohydrates), antibacterial agents (e.g., benzoic acid, casein, gentamycin), defoamers (e.g., polydimethyl baby pay (polydimethylsilozone)), preservatives (e.g., sulfur Liu Li, 2-phenoxyethanol, EDTA), polymeric stabilizers and viscosity modifiers (e.g., polyethylene of slightly associative paid, poloxamer 488, methylcellulose cluster), W and co-solvent (e.g., glycerin, polyethylene glycol, ethanol).

[0230] 根据本发明的组合物包含与药学上可接受的赋形剂组合的本发明的合成纳米载体。 [0230] A synthetic nanocarriers of the present invention in combination with a pharmaceutically acceptable excipient The composition of the present invention. 可W使用常规药物制造和配制技术制造运些组合物,W实现有用的剂型。 W may be manufactured using conventional pharmaceutical formulating techniques and manufacturing operation these compositions, W useful dosage forms. 适合用于实践本发明的技术可W在W下文献中找到:爱德华· L保罗化dward L.Paul),维克多· A.Atiem〇-〇beng(Victor A.Atiem〇-〇beng)W及苏珊娜· M.Kresta(Suzanne M.Kresta)编著的工业混合手册:科学与实践(Handbook of Industrial Mixing:Science and Practice) ,2004约翰威利国际出版公司(John Wiley&Sons,Inc.);和奥斯下· ME (Μ.E.Auten)编著的制药学:剂型设计科学(Pha;rmaceutics:The Science of Dosage 化!·!!! Desi即),第2版,2001,丘吉尔利文斯通出版社(化urchi 11 Livingstone)。 Suitable techniques useful in the practice of the present invention can be found in W W in the literature: · L Paul Edward of dward L.Paul), Victor A.Atiem〇-〇beng (Victor A.Atiem〇-〇beng) W and Susanna · M.Kresta (Suzanne M.Kresta) edited by industrial mix Handbook: Science and practice (Handbook of industrial mixing: Science and practice), 2004 John Wiley Publishing company (John Wiley & Sons, Inc.); and Oscar under · ME (Μ.E.Auten) compiled the pharmacy: dosage design Science (Pha; rmaceutics: the Science of dosage of · !!! Desi That!), 2nd edition, 2001, Churchill Livingstone Press ( of urchi 11 Livingstone). 在一个实施例中,将本发明的合成纳米载体与防腐剂一起悬浮在注射用无菌盐水溶液中。 In one embodiment, suspended in sterile saline injectable aqueous medium together with a preservative synthetic nanocarriers of the present invention.

[0231] 应当理解可W按任何适合的方式制造本发明的运些组合物,并且本发明绝不局限于可W使用在此描述的方法所产生的组合物。 [0231] It should be understood that the present invention may be manufactured W is transported in any suitable manner these compositions, and the present invention is not limited to W may be used in the compositions described herein methods produced. 适当方法的选择可能需要注意被结合的特定部分的特性。 The method may select appropriate characteristics of a particular portion of the note to be bound.

[0232] 在一些实施例中,本发明的合成纳米载体是在无菌条件下制造或最后进行灭菌。 [0232] In some embodiments, synthetic nanocarriers of the present invention is to produce final or sterilized under aseptic conditions. 运可W确保所得的组合物是无菌的并且是非感染性的,因此当与非无菌的组合物相比时提高了安全性。 W may be transported to ensure that the resulting composition is sterile and non-infectious, thus improving safety when compared to non-sterile compositions. 运提供了有价值的安全措施,尤其是当接受合成纳米载体的受试者具有免疫缺陷、正在遭受感染、和/或易受感染时。 It provides valuable transport security measures, especially when synthetic nanocarriers accepted subject having immune deficiency, are suffering from infection, and / or susceptible to infection. 在一些实施例中,取决于配制策略,可W将本发明的合成纳米载体冻干并且储存在悬浮液中或呈冻干粉末形式,持续延长的时间而无活性损失。 In some embodiments, depending on the formulation strategies, W may be synthetic nanocarriers of the present invention is lyophilized and stored in suspension or in a lyophilized powder form, for an extended period of time without loss of activity.

[0233] 可W通过各种途径给予本发明的运些组合物,运些途径包括但不限于皮下、鼻内、 静脉内、腹膜内、肌肉内、透粘膜、透粘膜、舌下、直肠、眼睛、肺部、皮肤内、经皮肤、经皮或真皮内或运些途径的组合。 [0233] W may be dosed some compositions of the invention by various routes, some transport routes including but not limited to subcutaneous, intranasal, intravenous, intraperitoneal, intramuscular, transmucosal, transmucosal, sublingual, rectal, eyes, lungs, skin, transdermal, intradermal or dermal or a combination of these transport pathways. 给予途径还包括通过吸入或肺气溶胶来进行的给予。 Route of administration further comprises administration to the lung by inhalation or aerosol. 用于制备气溶胶递送系统的技术是本领域技术人员众所周知的(参见,例如夏拉(Sciarra)和库替(Cutie ),。气溶胶(AerosolS ),"雷明顿的药物科学(Remington ' S 化armaceutical Sciences),第18版,1990,第1694-1712页;该文献通过引用结合在此)。 Techniques for preparing aerosol delivery systems are well known to those skilled in the art (see, e.g. Chellah (, Sciarra) and libraries for (Cutie) ,. aerosol (AerosolS), "Remington's Pharmaceutical Sciences (Remington 'S of armaceutical Sciences), 18th Edition, 1990, pp. 1694-1712; which is incorporated by reference herein).

[0234] 作为本发明的基于细胞的治疗而提供的可移植的移植物或治疗性蛋白可W通过肠胃外、动脉内注射、鼻内或静脉内给予或通过注射至淋己结或眼前房或通过局部给予至感兴趣的器官或组织而进行给予。 [0234] As portable graft therapeutic protein or cell-based therapy provided by the present invention may be parenterally W, intraarterial injection, intranasal administration, or hexyl, or to lymph nodes or anterior chamber or by intravenous injection and it is administered by topical administration to the tissue or organ of interest. 可W通过皮下、銷内、屯、室内、肌肉内、腹膜内、冠状动脉内、膜腺内、肝内或支气管注射来进行给予。 W may be by subcutaneous, pin, Tun, intraventricular, intramuscular, intraperitoneal, intracoronary, the film glands, intrahepatic injection or bronchial be administered.

[0235] 本发明的运些组合物可W按有效的量给予,如在此其他地方描述的有效的量。 [0235] These compositions of the present invention, operation may be administered in an amount effective by W, an effective amount as described elsewhere herein. 根据本发明,剂型的剂量含有不同量的多个群体的合成纳米载体和/或不同量的抗原和/或免疫抑制剂。 According to the present invention, dosage forms containing the synthetic nanocarriers plurality of groups of different amounts and / or different amounts of antigen and / or an immunosuppressive agent. 本发明的剂型中存在的合成纳米载体和/或抗原和/或免疫抑制剂的量可w根据运些抗原和/或免疫抑制剂的性质、有待完成的治疗益处、W及其他此类参数而改变。 Or a dosage form of the present invention in the presence of synthetic nanocarriers and / or antigen and / or immunosuppressive agents may be transported according to the nature of these antigens w and / or an immunosuppressant, a therapeutic benefit to be done, W, and other such parameters change. 在多个实施例中,可W进行剂量范围研究,W建立有待存在于该剂型中的合成纳米载体群体的最佳治疗量W及抗原和/或免疫抑制剂的量。 In various embodiments, the dose range studied W, W to be present in an amount to establish optimal therapeutic amount W of the dosage form the antigen and synthetic nanocarriers groups and / or immunosuppressants. 在多个实施例中,运些合成纳米载体和/或运些抗原和/或免疫抑制剂是W-定的量存在于该剂型中,当向受试者给予时,该量有效于产生针对运些抗原的致耐受性免疫应答。 In various embodiments, some synthetic nanocarriers transport and / or transport of these antigens and / or immunosuppressive agents are present in an amount W- given the dosage form, when administered to a subject in an amount effective for generating these antigens transport tolerogenic immune response. 在受试者中使用常规剂量范围研究和技术,也许有可能确定对产生致耐受性免疫应答有效的抗原和/或免疫抑制剂的量。 Using conventional techniques and dose ranging study in a subject, it may be possible to determine the resistance to an amount effective immunization response induced by an antigen and / or an immunosuppressive agent. 可W在多种频率下给予本发明的剂型。 W may be administered in the dosage forms of the present invention at a variety of frequencies. 在一个优选的实施例中,至少一次给予该剂型足W产生药理学上相关的应答。 In a preferred embodiment, at least one administration of the dosage form sufficient to produce W pharmacologically related responses. 在多个更优选的实施例中,利用至少两次给予、至少Ξ次给予、或至少四次给予该剂型来确保药理学上相关的应答。 In a more preferred embodiment a plurality of embodiments, given with at least two, at least Ξ administered once, or at least four times to ensure administration of the dosage form of a pharmacologically relevant response.

[0236] 可W在疾病、素乱或病症发作之前启动运些发明组合物的预防性给予,或可W在素乱、素乱或病症建立之后启动治疗性给予。 [0236] W may be started before the disease, disorder or condition onset prime transport these compositions of the present invention administered prophylactically, or in prime W disorder, administration of a therapeutic after start hormone disorder or condition established.

[0237] 在一些实施例中,例如在给予一种治疗性蛋白或可移植的移植物或暴露于一种过敏原之前进行合成纳米载体的给予。 [0237] In some embodiments, for example, administration of a therapeutic protein or implantable graft or an allergen is administered prior to exposure to the synthesis nanocarriers. 在多个示例性的实施例中,在给予一种治疗性蛋白或可移植的移植物或暴露于一种过敏原之前包括但不限于30天、25天、20天、15天、14天、13 天、12天、11天、10天、9天、8天、7天、6天、5天、4天、3天、2天、1天、或0天给予合成纳米载体一次或多次。 In an exemplary embodiment a plurality of, the administration of a therapeutic protein or implantable graft or prior to exposure to an allergen including but not limited to 30 days, 25 days, 20 days, 15 days, 14 days, 13 days, 12 days, 11 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or synthetic nanocarriers 0 days administered one or more times . 另外或可替代地,可W在给予一种治疗性蛋白或可移植的移植物或暴露于一种过敏原之后向受试者给予合成纳米载体。 Additionally or alternatively, W may be the administration of a therapeutic protein or implantable graft or after exposure to an allergen administered to a subject synthetic nanocarriers. 在多个示例性的实施例中,在给予一种治疗性蛋白或可移植的移植物或暴露于一种过敏原之后包括但不限于1天、2天、3天、4天、5天、6天、7 天、8天、9天、10天、11天、12天、13天、14天、15天、20天、25天、30天等等而给予合成纳米载体一次或多次。 In an exemplary embodiment a plurality of, the administration of a therapeutic protein or implantable graft or after exposure to an allergen including but not limited to 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 20 days, 25 days, 30 days, etc. and synthetic nanocarriers administered one or more times.

[0238] 在一些实施例中,在初始给予已经在受试者中产生一种致耐受性应答之后向该受试者给予维持剂量(例如,在此提供的合成纳米载体组合物的维持剂量),例如W便维持在初始剂量之后所达到的致耐受性作用、W便防止该受试者中的所不希望的免疫应答、或W 便防止该受试者变为处于经历所不希望的免疫应答或所不希望的水平的免疫应答风险中的受试者。 [0238] In some embodiments, the initial administration has been generated (e.g., maintenance doses provided herein are synthetic nanocarriers compositions tolerogenic response after one kind of maintenance dose is administered to the subject in a subject ), such as W tolerance effect will be maintained after the initial dose reached actuator, W will prevent an immune response in the subject is undesirable, will prevent the subject or W is subjected to undesirably becomes immune immune response or undesirable level of risk responses in subjects. 在一些实施例中,该维持剂量是与该受试者接受的初始剂量相同的剂量。 In some embodiments, the maintenance dose is the same initial doses received with the subject. 在一些实施例中,该维持剂量是比初始剂量更低的剂量。 In some embodiments, the maintenance dose is a dose lower than the initial dose. 例如,在一些实施例中,该维持剂量是约3/4、约2/3、约1/2、约1/3、约1/4、约1/8、约1/10、约1/20、约1/25、约1/50、约1/100、约1/1, 000、约1/10,000、约1/100,000、或约1/1,000,000(重量/重量)的初始剂量。 For example, in some embodiments, the maintenance dose is about 3/4, about 2/3, about 1/2, about 1/3, about 1/4, about 1/8, about 1/10, about 1 / 20, about 1/25, about 1/50, about 1/100 to about 1/1, 000, from about 1 / 10,000, to about 1 / 100,000, or from about 1 / 000,000 (weight / weight) of the initial dose.

[0239] 出于免疫抑制的目的,在此描述的运些组合物和方法可W用来诱导或增强一种致耐受性免疫应答和/或抑制、调节、引导或重定向所不希望的免疫应答。 [0239] For the purposes of immunosuppressive, transport these compositions and methods described herein may be used to induce or enhance one W tolerogenic immune response and / or inhibiting, regulating, or redirect the guide undesirable the immune response. 在此描述的运些组合物和方法可W用于诊断、预防和/或治疗其中免疫抑制(例如,致耐受性免疫应答)将会赋予治疗益处的疾病、素乱或病症。 These compositions and methods of operation described herein may be W for the diagnosis, prevention and / or treatment of diseases where immunosuppression (e.g., tolerogenic immune response) will confer a therapeutic benefit of a disease, disorder or condition factors. 运类疾病、素乱或病症包括炎性疾病、自身免疫性疾病、过敏、器官或组织排斥W及移植物抗宿主病。 Yun diseases, hormone disorder includes an inflammatory disease or disorder, an autoimmune disease, allergy, organ or tissue rejection W and graft versus host disease. 在此描述的运些组合物和方法还可W用于已经经历或将要经历移植的受试者。 These compositions and methods of operation described herein may be used subject W that has undergone or will undergo transplantation. 在此描述的运些组合物和方法还可W用于已经接受、正在接受或将要接受一种治疗性蛋白的受试者,针对该治疗性蛋白,该受试者已经产生或预期会产生一种所不希望的免疫应答。 These compositions and methods of operation described herein may be used for W has received, is being or will be subject receiving a therapeutic protein, the therapeutic protein against which the subject has been generated or expected to produce a kinds of undesired immune responses.

[0240] 自身免疫性疾病包括但不限于类风湿性关节炎、系统性硬化、免疫介导的或I型糖尿病、炎性肠病(例如,克罗恩病或溃瘍性结肠炎)、系统性红斑狼疮、银屑病、硬皮病、自身免疫性甲状腺疾病、斑秀、格雷夫病、格林-己利综合征、乳糜泻、干燥综合征、风湿热、胃炎、 自身免疫性萎缩性胃炎、自身免疫性肝炎、膜岛炎、卵巢炎、睾丸炎、葡萄膜炎、晶状体源性葡萄膜炎、重症肌无力、原发性粘液性水肿、恶性贫血、自身免疫性溶血性贫血、阿狄森病、 硬皮病、古德帕斯彻氏综合征、肾炎(例如,肾小球肾炎)、银屑病、寻常性天瘤疮、类天瘤疮、 交感性眼炎、特发性血小板减少性紫齋、特发性白细胞减少症(feucopenia)、韦格纳肉芽肿病W及多/皮肌炎。 [0240] Autoimmune diseases include, but are not limited to, rheumatoid arthritis, systemic sclerosis, immune-mediated or type I diabetes, inflammatory bowel disease (e.g., Crohn's disease or ulcerative colitis), systemic lupus erythematosus, psoriasis, scleroderma, autoimmune thyroid disease, plaques show, Graves disease, Guillain - has benefits syndrome, celiac disease, Sjogren's syndrome, rheumatic fever, gastritis, autoimmune atrophic gastritis, autoimmune hepatitis, inflammatory film islands, oophoritis, orchitis, uveitis, lens endogenous uveitis, myasthenia gravis, primary myxedema, pernicious anemia, autoimmune hemolytic anemia, Addison disease, scleroderma, Goodpasture's syndrome, nephritis (e.g., glomerulonephritis), psoriasis, acne vulgaris day tumors, tumor type sores days, sympathetic ophthalmia, idiopathic thrombocytopenia fast violet, idiopathic leukopenia (feucopenia), Wegener's granulomatosis and multiple W / dermatomyositis.

[0241] -些另外的示例性的自身免疫性疾病、相关联的自身抗原、W及自身抗体(它们预期会在本发明中使用)在W下表1中描述: [0241] - some further exemplary autoimmune diseases associated autoantigens, W, and autoantibodies (which is expected to be used in the present invention) is described in Table 1 W:

[0242] [0242]

Figure CN105999295AD00481

Figure CN105999295AD00491

Figure CN105999295AD00501

Figure CN105999295AD00511

[0246]炎性疾病包括但不限于:阿尔茨海默病、强直性脊柱炎、关节炎、哮喘、动脉粥样硬化、白塞病、慢性炎性脱髓銷性多发性神经根神经病、克罗恩病、结肠炎、囊性纤维化、皮炎、 憩室炎、肝炎、肠易激综合征(IBS )、红斑狼疮、肌营养不良症、肾炎、帕金森病、带状瘤疹和溃瘍性结肠炎。 [0246] Inflammatory diseases include, but are not limited to: Alzheimer's disease, ankylosing spondylitis, arthritis, asthma, atherosclerosis, Behcet's disease, chronic inflammatory demyelinating polyradiculoneuropathy exemplary pin, g Ron's disease, colitis, cystic fibrosis, dermatitis, diverticulitis, hepatitis, irritable bowel syndrome (IBS), lupus erythematosus, muscular dystrophy, nephritis, Parkinson's disease, and ulcerative colitis strip tumors rash inflammation. 炎性疾病还包括,例如,屯、血管疾病、慢性阻塞性肺疾病(C0PD)、支气管扩张、慢性胆囊炎、结核病、桥本氏甲状腺炎、败血病、结节病、秒肺及其他尘肺、W及伤口中的植入异体、但都不局限于此。 Inflammatory diseases also include, e.g., Tun, vascular diseases, chronic obstructive pulmonary disease (C0PD), bronchiectasis, chronic cholecystitis, tuberculosis, Hashimoto's thyroiditis, sepsis, sarcoidosis, lung and other pneumoconiosis seconds , W, and the wound allografted, but not limited thereto. 如在此所使用,术语"败血症"是指一种公认的与针对微生物侵袭的宿主系统性炎症应答相关联的临床综合征。 As used herein, the term "sepsis" refers to a recognized response associated with microbial attack against a host of clinical syndromes of systemic inflammation. 在此所使用的术语"败血症"是指典型地信号为发热或低体溫、屯、动过速w及呼吸急促的一种病症,并且在严重的情况下可进展为低血压、器官功能失常,甚至死亡。 As used herein, the term "sepsis" refers to the signal is typically fever or hypothermia, village, tachycardia and tachypnea w of a condition, and can progress to hypotension, organ dysfunction in severe cases, even death.

[0247]在一些实施例中,炎性疾病是非自身免疫性炎性肠病、手术后粘连、冠状动脉疾病、肝纤维化、急性呼吸窘迫综合征、急性炎性膜腺炎、内窥镜逆行胆管膜造影术引起的膜腺炎、烧伤、冠状动脉粥样硬化、脑和外周动脉、阔尾炎、胆囊炎、憩室炎、内脏纤维化病症、 伤口愈合、皮肤癒痕形成病症(癒痕瘤、化脈性汗腺炎)、肉芽肿性病症(结节病、原发性胆汁性肝硬变)、哮喘、坏痘性脈皮病、Sweet氏综合征、白塞病、原发性硬化性胆管炎或脈肿。 [0247] In some embodiments, non-inflammatory diseases autoimmune inflammatory bowel disease, postoperative adhesion, coronary artery disease, hepatic fibrosis, acute respiratory distress syndrome, acute inflammatory pancreatitis, endoscopic retrograde angiography films biliary pancreatitis caused by burns, coronary atherosclerosis, cerebral and peripheral arteries, broad tail, cholecystitis, diverticulitis, visceral fibrotic disorders, wound healing, skin disorders formed more marks (marks more tumor , hidradenitis of pulse), granulomatous disorders (sarcoidosis, primary biliary cirrhosis), asthma, bad veins pox skin disease, Sweet's syndrome, Behcet's disease, primary sclerosing cholangitis or swollen veins. 在一些优选的实施例中,炎性疾病是炎性肠病(例如,克罗恩病或溃瘍性结肠炎)。 In some preferred embodiments, the inflammatory disease is inflammatory bowel disease (e.g., Crohn's disease or ulcerative colitis).

[024引在其他实施例中,炎性疾病是一种自身免疫性疾病。 [024] In other embodiments cited embodiment, the inflammatory disease is an autoimmune disease. 在一些实施例中自身免疫性疾病是类风湿性关节炎、风湿热、溃瘍性结肠炎、克罗恩病、自身免疫性炎性肠病、膜岛素依赖型糖尿病、糖尿病、青少年糖尿病、自发性自身免疫性糖尿病、胃炎、自身免疫性萎缩性胃炎、自身免疫性肝炎、甲状腺炎、桥本氏甲状腺炎、膜岛炎、卵巢炎、睾丸炎、葡萄膜炎、晶状体源性葡萄膜炎、系统性硬化、重症肌无力、原发性粘液性水肿、甲状腺功能亢进、恶性贫血、自身免疫性溶血性贫血、阿狄森病、强直性脊柱炎、结节病、硬皮病、肺出血肾炎综合征、 格林-己利综合征、格雷夫病、肾小球肾炎、银屑病、寻常性天瘤疮、类天瘤疮、湿疹、大瘤性类天瘤疮、交感性眼炎、特发性血小板减少性紫齋、特发性白细胞减少症(feucopenia)、干燥综合征、系统性硬化、韦格纳肉芽肿、多肌炎/皮肌炎、原发性胆汁性肝硬变、 In some embodiments, the autoimmune disease is rheumatoid arthritis, rheumatic fever, ulcerative colitis, Crohn's disease, autoimmune inflammatory bowel disease, Insulin dependent diabetes film, diabetes, juvenile diabetes, spontaneous autoimmune diabetes, gastritis, autoimmune atrophic gastritis, autoimmune hepatitis, thyroiditis, Hashimoto's thyroiditis, inflammatory film islands, oophoritis, orchitis, uveitis, endogenous uveitis lens, systemic sclerosis, myasthenia gravis, primary myxedema, hyperthyroidism, pernicious anemia, autoimmune hemolytic anemia, Addison's disease, ankylosing spondylitis, sarcoidosis, scleroderma, Goodpasture syndrome, Green - has benefits syndrome, Graves disease, glomerulonephritis, psoriasis, acne vulgaris day tumors, tumor-day class sores, eczema, tumors of the large class of tumor-day sore, sympathetic ophthalmia, special idiopathic thrombocytopenic purpura fasting, idiopathic leukopenia (feucopenia), Sjogren's syndrome, systemic sclerosis, Wegener's granulomatosis, polymyositis / dermatomyositis, primary biliary cirrhosis, 发性硬化性胆管炎、红斑狼疮或系统性红斑狼疮。 Cholangitis sclerosis, lupus or systemic lupus erythematosus.

[0249]移植物抗宿主病(GV皿)是多能细胞(例如,干细胞)或骨髓移植之后可W发生的并发症,其中新移植的材料对移植受体身体产生攻击。 [0249] graft-versus-host disease (GV dish) are pluripotent cells (e.g., stem cells), or after bone marrow transplantation complications may occur W, wherein the graft material to produce a new attack on the transplant recipient body. 在一些实例中,GVHD在输血后发生。 In some examples, GVHD occurs after transfusion. 移植物抗宿主病可W分为急性和慢性形式。 Graft versus host disease W can be divided into acute and chronic forms. 急性或暴发形式的疾病(aGVHD)通常在移植后头100天观察到,并且由于相关的发病率和死亡率它对移植物是一项重大挑战。 Acute or fulminant forms of the disease (of aGVHD) transplant typically observed during the first 100 days, and since the morbidity and mortality associated with graft it is a major challenge. 慢性形式的移植物抗宿主病(cGVHD)通常发生在100天之后。 Chronic forms of graft-versus-host disease (of cGVHD) typically occurs after 100 days. cGVHD的中度至重度情况的出现有害地影响长期存活。 cGVHD appearance of moderate to severe cases adversely affect long-term survival.

[0巧日]实例 [0 Qiao day] examples

[0251]实例1:具有或不具有卵白蛋白肤(323-339)的带有偶联的雷帕霉素的合成纳米载体的免疫应答[0况]材料 [0251] Example 1: Synthesis of immune nanocarriers rapamycin coupled with ovalbumin with or without skin (323-339) response [0 conditions] Material

[0巧3] 从己亨美洲公司(Bachem Americas Inc.,柏市街化ashiwa Sl:reet)3132号,托伦斯(Torrance)加利福尼亚州(CA)90505;区号#4065609)购买卵白蛋白肤323-339,即一种已知为卵白蛋白的T和B细胞表位的17氨基酸肤。 [0 Qiao 3] has been from Hang Americas (Bachem Americas Inc., cypress urbanization ashiwa Sl: reet) No. 3132, Torrance (Torrance), California (CA) 90505; area code # 4065609) purchased ovalbumin skin 323- 339, i.e., one known as the 17 amino acid skin ovalbumin T and B cell epitopes. 从TSZ化学品公司(TSZ畑EM,威尔逊街(Wilson Street)185 号,弗雷明汉(Framin 曲am),马萨诸塞州(MA)01702;产品目录#31017) 购买雷帕霉素。 Buy rapamycin; from the TSZ Chemical Company (Catalog # 31017 TSZ Hata EM, Wilson Street (Wilson Street) No. 185, Framingham (Framin song am), Massachusetts (MA) 01702). 从SurModics制药公司(SurModics Pharmaceuticals,汤姆马下路(Tom Madin Drive)756号,伯明翰(Birmin曲am),亚拉己马州(AL)35211;产品代码752加LG 7A) 购买丙交醋:乙交醋比为3:1并且特性粘度为0.75化/g的化GA。 SurModics from pharmaceutical companies (SurModics Pharmaceuticals,) 756 Tom Muma down the road (Tom Madin Drive, Birmingham (Birmin song am), Yala had Maryland (AL) 35211; product code 752 plus LG 7A) buy lactide vinegar: B cross vinegar ratio of 3: 1 and an intrinsic viscosity of 0.75 / g, of the GA. 从EMD化学品公司化MD Chemicals,产品编号1.41350.1001)购买聚乙締醇(85%-89%水解的)。 Polyvinyl alcohol for later association (85% -89% from the EMD Chemicals Inc. of MD Chemicals, Catalog No .: 1.41350.1001) hydrolysis).

[0254]溶液1:在稀释的盐酸水溶液中的卵白蛋白肤323-339@20mg/mL。 [0254] solution 1: aqueous solution of hydrochloric acid diluted skin ovalbumin 323-339 @ 20mg / mL. 通过在室溫下将卵白蛋白肤溶解在0.13M盐酸溶液中而制备该溶液。 The solution prepared in 0.13M hydrochloric acid solution by the skin of ovalbumin dissolved at room temperature.

[0巧5]溶液2:在二氯甲烧中的雷帕霉素@50mg/mL。 [Qiao 0 5] solution 2: Burning in dichloromethane rapamycin @ 50mg / mL. 通过将雷帕霉素溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by dissolving the pure rapamycin dichloromethane calcining.

[0巧6] 溶液3:在二氯甲烧中的PLGA@1 OOmg/mL。 [Qiao 0 6] Solution 3: In the burning of dichloromethane PLGA @ 1 OOmg / mL. 通过将PLGA溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by dissolving the PLGA in neat dichloromethane calcining.

[0257] 溶液4:在lOOmM的抑8的憐酸盐缓冲液中的聚乙締醇@50mg/mL。 [0257] Solution 4: pity in acetate buffer lOOmM t08 in the association of alcohol polyethylene @ 50mg / mL.

[0258] 用于制备含有雷帕霉素和卵白蛋白(323-339)的合成纳米载体的方法 The method [0258] for the preparation of synthetic nanocarriers containing rapamycin and ovalbumin (323-339) of

[0259] 首先制备第一油包水乳液。 [0259] First preparing a first water in oil emulsions. 通过将溶液1 (0.2mL)、溶液2(0.2mL)、W及溶液3 (l.OmL)合并在一个小的压力管中并且使用布兰森数字超声波仪(Branson Digital Sonifier)250在50%振幅下进行声处理40秒来制备Wl/01。 By the solution 1 (0.2mL), a solution of 2 (0.2mL), W and solution (l.OmL) 3 were combined in a small pressure tube and using Branson Digital Sonifier (Branson Digital Sonifier) ​​250 50% amplitude for 40 seconds sonication prepared Wl / 01. 然后通过将溶液4(3.0mL)与第一W1/01乳液合并、满旋10秒、并且使用布兰森数字超声波仪250在30%振幅下进行声处理60秒来制备第二乳液(W1/01/W2)。 The solution was then passed through 4 (3.0mL) and the first W1 / 01 emulsion were combined, spin full 10 seconds, and using the Branson Digital Sonifier 250 acoustically at 30% amplitude for 60 seconds to prepare a second emulsion (W1 / 01 / W2).

[0260] 将该W1/01/W2乳液添加到含有70mM的pH如溝酸盐缓冲溶液(30mL)的烧杯中并且在室溫下揽拌2小时W允许二氯甲烧蒸发并且允许合成纳米载体形成。 [0260] The added W1 / 01 / W2 emulsion to the beaker containing the groove as pH 70mM acetate buffer solution (30mL) and embrace the W for 2 hours to allow dichloromethane and allowed to burn synthetic nanocarriers evaporated at room temperature form. 通过W下方式来洗涂一部分合成纳米载体:将合成纳米载体悬浮液转移至一个离屯、管中,并且在21,000Xg和4°C下离屯、一小时、去除上清液、并且将沉淀再悬浮于憐酸盐缓冲盐水中。 By way of a portion of the W-coated to wash the synthetic nanocarriers: synthetic nanocarrier suspension was transferred to a village from the tube, and 21,000Xg at 4 ° C and from the village, one hour, the supernatant was removed, and the Rei acid pellet was resuspended in buffered saline. 重复该洗涂程序, 并且将沉淀再悬浮于憐酸盐缓冲盐水中W用于约lOmg/mL的最终的合成纳米载体分散液。 The wash-coating procedure was repeated, and the pellet was resuspended in acetate buffer saline W pity for final synthetic nanocarriers of about lOmg / mL dispersion. [0261 ]通过HPLC分析来确定在合成纳米载体中的肤和雷帕霉素的量。 [0261] determined by HPLC analysis and the amount of skin in the synthesis of rapamycin nanocarrier. 通过重量法来确定每mL悬浮液的总的干燥合成纳米载体质量。 To determine the total mass of the dried synthetic nanocarriers per mL of suspension was determined by the weight method.

[0262] 用于含有雷帕霉素的合成纳米载体的方法 The method [0262] for the synthesis of nanocarrier comprising rapamycin

[0263] 首先制备第一油包水乳液。 [0263] First preparing a first water in oil emulsions. 通过将0.13M盐酸溶液(0.2mL)、溶液2(0.2mL)、W及溶液3(1.OmL)合并在一个小的压力管中并且使用布兰森数字超声波仪250在50%振幅下进行声处理40秒来制备W1/01。 By 0.13M hydrochloric acid solution (0.2mL), a solution of 2 (0.2mL), W, and a solution of 3 (1.OmL) were combined in a small pressure tube and using Branson Digital Sonifier 250 at 50% amplitude sonicated for 40 seconds to prepare W1 / 01. 然后通过将溶液4(3.0mL)与第一W1/01乳液合并、满旋10秒、并且使用布兰森数字超声波仪250在30%振幅下进行声处理60秒来制备第二乳液(W1/01/W2)。 The solution was then passed through 4 (3.0mL) and the first W1 / 01 emulsion were combined, spin full 10 seconds, and using the Branson Digital Sonifier 250 acoustically at 30% amplitude for 60 seconds to prepare a second emulsion (W1 / 01 / W2).

[0264] 将该W1/01/W2乳液添加到含有70mM的pH如溝酸盐缓冲溶液(30mL)的烧杯中并且在室溫下揽拌2小时W允许二氯甲烧蒸发并且允许合成纳米载体形成。 [0264] The added W1 / 01 / W2 emulsion to the beaker containing the groove as pH 70mM acetate buffer solution (30mL) and embrace the W for 2 hours to allow dichloromethane and allowed to burn synthetic nanocarriers evaporated at room temperature form. 通过W下方式来洗涂一部分合成纳米载体:将合成纳米载体悬浮液转移至一个离屯、管中,并且在21,000Xg和4°C下离屯、一小时、去除上清液、并且将沉淀再悬浮于憐酸盐缓冲盐水中。 By way of a portion of the W-coated to wash the synthetic nanocarriers: synthetic nanocarrier suspension was transferred to a village from the tube, and 21,000Xg at 4 ° C and from the village, one hour, the supernatant was removed, and the Rei acid pellet was resuspended in buffered saline. 重复该洗涂程序, 并且将沉淀再悬浮于憐酸盐缓冲盐水中W用于约lOmg/mL的最终的合成纳米载体分散液。 The wash-coating procedure was repeated, and the pellet was resuspended in acetate buffer saline W pity for final synthetic nanocarriers of about lOmg / mL dispersion.

[0265] 通过HPLC分析来确定在合成纳米载体中的雷帕霉素的量。 [0265] to determine the amount of rapamycin in the synthesis nanocarriers by HPLC analysis. 通过重量法来确定每mL 悬浮液的总的干燥合成纳米载体质量。 To determine the total mass of the dried synthetic nanocarriers per mL of suspension was determined by the weight method.

[0266] 用于测量雷帕霉素负载量的方法 [0266] A method for measuring the load of rapamycin

[0267] 收集近似3mg的合成纳米载体,并且离屯、W使上清液与合成纳米载体沉淀分离。 [0267] collected approximately 3mg synthetic nanocarriers, and from the village, W precipitate separated from the supernatant synthetic nanocarriers. 向该沉淀添加乙腊,并且对样品进行声处理并且离屯、W去除任何不溶的材料。 Acetate was added to this precipitate wax, and the sample sonicated and from the village, W remove any insoluble material. 将该上清液和沉淀注射在RP-HPLC上,并且在278nm处读取吸光度。 The supernatant and pellet injection on RP-HPLC, and the absorbance was read at 278nm. 该沉淀中发现的yg用来计算%包载(负载),在上清液和沉淀中的yg用来计算回收的总yg。 The precipitate was found yg used to calculate the% entrapped (load), the supernatant and pellet yg used to calculate the total yg recovered.

[0268] 用于测量卵白蛋白(323-339)负载量的方法 The method [0268] for measuring (323-339) loading ovalbumin

[0269] 收集近似3mg的合成纳米载体,并且离屯、W使上清液与合成纳米载体沉淀分离。 [0269] collected approximately 3mg synthetic nanocarriers, and from the village, W precipitate separated from the supernatant synthetic nanocarriers. 向该沉淀添加Ξ氣乙醇并且对样品进行声处理W溶解该聚合物,添加〇.2%Ξ氣乙酸并且对样品进行声处理并且然后离屯、W去除任何不溶的材料。 To this ethanol precipitate was added a Cascade gas samples were sonicated and W dissolve the polymer, is added 〇.2% Ξ acid gas and the sample was sonicated and then from the village, W remove any insoluble material. 将该上清液和沉淀注射在RP-HPLC 上,并且在215nm处读取吸光度。 The supernatant and pellet injection on RP-HPLC, and the absorbance was read at 215nm. 该沉淀中发现的yg用来计算%包载(负载),在上清液和沉淀中的yg用来计算回收的总yg。 The precipitate was found yg used to calculate the% entrapped (load), the supernatant and pellet yg used to calculate the total yg recovered.

[0270] 在化eg细胞发育上的抗原特异性致耐受性树突状细胞(Tdc)活性 [0270] antigens on specific cell development eg of tolerogenic dendritic cells (Tdc) activity

[0271] 该测定包括使用具有对免疫显性卵白蛋白(323-339)特异的转基因T细胞受体的0ΤΠ 小鼠。 [0271] The assay involves the use of mice having 0ΤΠ immunodominant ovalbumin (323-339) specific transgenic T cell receptor. 为了产生抗原特异性tDC,将CD1 lc+脾脏细胞分离,并且在化g/ml或无抗原下在体外添加卵白蛋白(323-339)肤。 To generate antigen-specific TDC, the spleen cells isolated CD1 lc +, and adding ovalbumin (323-339) in vitro in the skin of g / ml or no antigen. 然后将可溶的或纳米载体封装的雷帕霉素添加到DC中保持2小时,然后将运些DC充分洗涂W从培养物中去除游离雷帕霉素。 Then soluble or encapsulated nanocarriers rapamycin added to the DC for 2 hours, then transport these DC were washed extensively to remove free coating W rapamycin from the culture. 将纯化的应答CD4+ CD25-细胞从0ΤΠ 小鼠分离并且W10:1的T与DC比添加到tDC中。 The response of purified CD4 + CD25- cells from the mouse and 0ΤΠ W10: T 1 ratio of DC and added to the tDC. 然后将tDC与0ΤΠΤ-细胞的混合物培养4-5天,并且通过如图1所示的流式细胞术来分析化eg细胞(CD4+CD25hi曲化xP3 + )的频率。 The mixture was then tDC 0ΤΠΤ- cells cultured for 4-5 days, and analyzed the frequency of eg cells (CD4 + CD25hi curvature of xP3 +) by flow cytometry as shown in Fig. 基于同种型对照选择区域。 Isotype control based on the selected area. 陶]实例2:具有偶联的布洛芬的介孔二氧化娃纳米颗粒(预示的) Tao] Example 2: Ibuprofen coupled with baby mesoporous silica nanoparticles (Prophetic)

[0273] 介孔Si〇2纳米颗粒核屯、通过溶胶-凝胶法来产生。 [0273] Mesoporous Si〇2 Tun nanoparticle core, by the sol - gel method to produce. 将十六烷基Ξ甲基漠化锭(CTAB) (0.5g)溶解在去离子水(500mL)中,并且然后将2M的化0H水溶液(3.5血)添加到CTAB 溶液中。 The methyl hexadecyl Ξ desertification ingot (CTAB) (0.5g) was dissolved in deionized water (500 mL), and then a 2M aqueous solution of 0H (3.5 of blood) was added to the CTAB solution. 将该溶液揽拌30分钟,并且然后向该溶液中添加四乙氧基硅烷(TE0S) (2.5mL)。 The solution was stirred for 30 minutes embrace, and then tetraethoxysilane (TE0S) (2.5mL) to this solution. 在80°C的溫度下将生成的凝胶揽拌3小时。 At a temperature to 80 ° C and the resulting gel embrace for 3 hours. 通过过滤捕获形成的白色沉淀,随后用去离子水洗涂并且在室溫下干燥。 By capturing the white precipitate formed was filtered, washed with deionized water and then coated and dried at room temperature. 然后通过悬浮于HC1的乙醇溶液中过夜而从颗粒中提取出残留的表面活性剂。 It is then extracted by suspension in a solution of HC1 in ethanol overnight from the particles of the residual surfactant. 将运些颗粒用乙醇洗涂、离屯、、并且在超声处理下再分散。 The transport of these particles are coated washed with ethanol, and then dispersed from the village ,, under sonication. 运个洗涂程序另外再重复两次。 Further transport a wash coating procedure was repeated twice.

[0274] 然后使用(3-氨基丙基)-Ξ乙氧基硅烷(APTMS)用氨基对Si化纳米颗粒进行功能化。 [0274] then (3-aminopropyl) -Ξ ethoxysilane (the APTMS) be functionalized with amino Si nanoparticles. 为此,将颗粒悬浮在乙醇(3〇1^)中,并且将4口了18(5化山添加到悬浮液中。允许该悬浮液在室溫下静置2小时,并且然后煮沸4小时,通过周期性地添加乙醇使体积保持恒定。残留的反应物通过五个离屯、洗涂和再分散于纯乙醇中的循环来去除。 For this purpose, the particles suspended in ethanol (3〇1 ^), and adding 4 of 18 (5 of mountain to the suspension. The suspension was allowed to stand at room temperature for 2 hours and then boiled for 4 hours by adding ethanol periodically to maintain a constant volume. the residue from the reaction was purified by five Tun, wash coating, and redispersed in pure ethanol to remove the loop.

[0275] 在一个单独的反应中,产生l-4nm直径的金种子。 [0275] In a separate reaction, to produce gold seed l 4nm-diameter. 在此反应中使用的所有的水首先被去离子并且然后从玻璃蒸馈。 All water used in this reaction is first distilled and then fed to an ion from the glass. 将水(45.5mL)添加到lOOmL圆底烧瓶中。 Water (45.5 mL) was added to a lOOmL round bottom flask. 在揽拌下,添加0.2M水性化0H( 1.5mL),随后添加氯化四(径甲基)鱗(THPC)的1 %水溶液(1. OmL)。 In embrace mix was added 0.2M aqueous based 0H (1.5mL), followed by the addition tetrachloride (meth diameter) scale (of THPC) 1% aqueous solution (1. OmL). 在添加THPC溶液之后两分钟,添加已经老化至少15分钟的氯金酸的lOmg/mL水溶液(2mL)。 lOmg / mL solution (2mL) two minutes after the addition was added solution THPC has been aged for at least 15 minutes chloroauric acid. 通过用水透析来纯化运些金种子。 These transport SEED purified by dialysis against water.

[0276] 为了形成核-壳纳米载体,将W上形成的氨基功能化的Si化纳米颗粒首先在室溫下与运些金种子混合2小时。 [0276] In order to form a core - shell nanocarriers, amino functionalized nanoparticles formed of Si on the W first at room temperature and transported more gold seeds mixed for 2 hours. 将金修饰的Si化颗粒通过离屯、来收集,并且与氯金酸和碳酸氨钟的水溶液混合来形成金外壳。 Si-modified gold particles by ion Tun, collected, and mixed with an aqueous solution of chloroauric acid and ammonium carbonate to form a gold bell housing. 然后将运些颗粒离屯、洗涂、并且再分散于水中。 These particles are then transported from the village, wash coating, and redispersed in water. 通过将运些颗粒悬浮于布洛芬钢的溶液(Img/L)中72小时来负载布洛芬。 Will be transported by these steel particles are suspended in a solution of ibuprofen (Img / L) for 72 hours to load ibuprofen. 然后将游离的布洛芬通过离屯、从颗粒中洗涂出并且再分散于水中。 Free ibuprofen is then isolated by Tun, from the coated particles are washed and re-dispersed in water. 脚7] 实例3:含有环抱霉素A的脂质体(预示的) Pin 7] Example 3: liposomes containing a cyclosporine A (Prophetic)

[027引运些脂质体使用薄膜水合来形成。 [027 cited these liposomes op formed using a thin film hydration. 将1,2-二栋桐酷-sn-甘油基-3-憐酸胆碱(DPPC)(32皿ol)、胆固醇(32皿ol)、W及环抱霉素A(6.4皿ol)溶解在纯氯仿(3血)中。 1,2 Kazuto two cool -sn- glycero-3-pity acid choline (of DPPC) (dish 32 ol), cholesterol (32 dish ol), W and cyclosporine A (6.4 dish OL) was dissolved in pure chloroform (blood) in. 将此脂质溶液添加到50mL的圆底烧瓶中,并且在60°C的溫度下在旋转蒸发器上蒸发溶剂。 This lipid solution was added to a 50mL round bottom flask, and the solvent was evaporated on a rotary evaporator at a temperature of 60 ° C. 烧瓶然后用氮气冲洗W去除残留溶剂。 W flask was then flushed with nitrogen to remove residual solvent. 将憐酸盐缓冲盐水(2mL)和五个玻璃珠添加到烧瓶中,并且通过在60°C下摇晃1小时来使脂质膜水合W形成悬浮液。 The pity-buffered saline (2mL) and five glass beads were added to the flask, and the lipid film is hydrated to by shaking at 60 ° C 1 hours W form a suspension. 将该悬浮液转移到一个小的压力管中并且在60°C下进行声处理,持续四个30秒脉冲循环,其中在每个脉冲之间有30秒的延迟。 The suspension was transferred to a small pressure tube and sonicated at 60 ° C, 30 seconds duration four pulse cycles, including a 30 second delay between each pulse. 然后使悬浮液在室溫下静置2小时,W允许完全水合。 The suspension is then allowed to stand at room temperature for 2 hours, W allow complete hydration. 离屯、洗涂脂质体,随后再悬浮于新鲜憐酸盐缓冲盐水中。 From Tun, wash coating the liposomes, then resuspended in fresh buffered saline pity salt. 陶]实例4:含有聚合物-雷帕霉素共辆物的聚合纳米载体(预示的) Tao] Example 4: Polymer comprising - co-rapamycin was polymerized units nanocarrier (Prophetic)

[0280] PLGA-雷帕霉素共辆物的制备: [0280] PLGA- rapamycin co vehicle was prepared:

[0281] 将具有酸端基的PLGA聚合物(752抓LG1A,酸值0.46mmol/g,Lakeshore生物材料公司(Xakeshore Biomaterials) ;5g,2.3mmol,1.0当量)溶解在30mL的二氯甲烧(DCM)中。 [0281] The PLGA polymer having acid end groups (752 grasping LG1A, an acid value of 0.46mmol / g, Lakeshore Biological Materials (Xakeshore Biomaterials); 5g, 2.3mmol, 1.0 eq) was dissolved in 30mL of dichloromethane burn ( DCM) in. 添加N,N-二环己基碳二亚胺(1.2当量,2.8mmol,0.57g),随后添加雷帕霉素(1.0当量, 2.3111111〇1,2.1旨)和4-二甲氨基化晚(0魁?)(2.0当量,4.6111111〇1,0.56旨)。 Was added N, N- dicyclohexyl carbodiimide (1.2 equiv., 2.8mmol, 0.57g), was added followed by rapamycin (1.0 equiv., 2.3111111〇1,2.1 purpose) and 4-dimethylaminopyridine LATE (0 Quebec?) (2.0 equiv., 4.6111111〇1,0.56 purpose). 将混合物在室溫下揽拌2天。 The mixture was stirred at room temperature for 2 embrace days. 然后过滤混合物W去除不溶的二环己脈。 The mixture was then filtered to remove insoluble dicyclohexyl W pulse. 将滤液浓缩至体积大约为lOmL并且添加到lOOmL的异丙醇(IPA)中W沉淀出PLGA-雷帕霉素共辆物。 The filtrate was concentrated to a volume of approximately lOmL lOOmL and added to isopropanol (IPA) in PLGA- W rapamycin co-precipitated units thereof. 将IPA层去除并且聚合物然后用50mL的IPA和50mL的甲基叔下酸(MTBE)洗涂。 The IPA layer was then removed and the acid polymer (MTBE) with methyl t-IPA 50mL and 50mL washcoat. 然后将聚合物在35°C下真空干燥2天,给出呈白色固体的PLGA-雷帕霉素(大约6.5g)。 The polymer was then vacuum dried for 2 days at 35 ° C, as a white solid PLGA- give rapamycin (about 6.5g).

[0282] 含有PLGA-雷帕霉素共辆物和卵白蛋白肤(323-339)的纳米载体的制备: [0282] PLGA- containing rapamycin was co vehicle skin and ovalbumin (323-339) nanocarriers were prepared:

[0283] 根据实例1中所述的程序如下制备含有PLGA-雷帕霉素的纳米载体: [0283] Preparation of a nanocarrier comprising PLGA- rapamycin according to the procedure described in Example 1 as follows:

[0284] 如下制备用于纳米载体形成的溶液: [0284] was prepared by forming a solution of the nanocarrier:

[0285] 溶液1:在稀释的盐酸水溶液中的卵白蛋白肤323-339@20mg/mL。 [0285] solution 1: aqueous solution of hydrochloric acid diluted skin ovalbumin 323-339 @ 20mg / mL. 通过在室溫下将卵白蛋白肤溶解在0.13M盐酸溶液中而制备该溶液。 The solution prepared in 0.13M hydrochloric acid solution by the skin of ovalbumin dissolved at room temperature. 溶液2:在二氯甲烧中的化GA-雷帕霉素@100mg/mL。 Solution 2: In the burning of dichloromethane GA- rapamycin @ 100mg / mL. 通过将PLGA-雷帕霉素溶解在纯二氯甲烧中而制备该溶液。 The solution is prepared by dissolving the pure rapamycin PLGA- dichloromethane calcining. 溶液3:在二氯甲烧中的PLA-PEG@ 1 OOmg/mL。 Solution 3: In the burning of dichloromethane PLA-PEG @ 1 OOmg / mL. 通过将化A-阳G溶解在纯二氯甲烧中而制备该溶液。 The solution is prepared by dissolving G of pure male A- dichloromethane calcining. 溶液4:在lOOmM的pH 8的憐酸盐缓冲液中的聚乙締醇@50mg/mL。 Solution 4: pity in acetate buffer pH lOOmM 8 in association polyvinyl alcohol @ 50mg / mL.

[0286] 首先制备第一油包水乳液。 [0286] First preparing a first water in oil emulsions. 通过将溶液1 (0.2mL)、溶液2 (0.75mL)、W及溶液3 (0.25mL)合并在一个小的压力管中并且使用布兰森数字超声波仪250在50%振幅下进行声处理40秒来制备W1/01。 By the solution 1 (0.2mL), a solution of 2 (0.75mL), W and a solution (0.25 mL of) 3 were combined in a small pressure tube and using Branson Digital Sonifier 250 at 50% amplitude sonication 40 seconds to prepare W1 / 01. 然后通过将溶液4(3.0mL)与第一W1/01乳液合并、满旋10秒、并且使用布兰森数字超声波仪250在30%振幅下进行声处理60秒来制备第二乳液(W1/01/W2)。 The solution was then passed through 4 (3.0mL) and the first W1 / 01 emulsion were combined, spin full 10 seconds, and using the Branson Digital Sonifier 250 acoustically at 30% amplitude for 60 seconds to prepare a second emulsion (W1 / 01 / W2). 将该W1/01/W2乳液添加到含有70mM的pH如溝酸盐缓冲溶液(30mL)的烧杯中并且在室溫下揽拌2小时W允许二氯甲烧蒸发并且允许纳米载体形成。 The W1 / 01 / W2 emulsion was added to a beaker containing grooves as pH 70mM acetate buffer solution (30mL) of W and embrace for 2 hours at room temperature to allow evaporation of dichloromethane and allowed to burn nanocarriers are formed. 通过W下方式来洗涂一部分纳米载体:将纳米载体悬浮液转移至一个离屯、管中,并且在75,600 Xg和4°C下离屯、35分钟、去除上清液、并且将沉淀再悬浮于憐酸盐缓冲盐水中。 By way of a portion of the W-coated to wash the nanocarrier: The nanocarrier suspension was transferred to a village from the tube, and at 75,600 Xg and 4 ° C from the village, 35 minutes, the supernatant was removed, and the pellet was resuspended pity salt in buffered saline. 重复该洗涂程序,并且将沉淀再悬浮于憐酸盐缓冲盐水中W用于约lOmg/mL的最终的纳米载体分散液。 The wash-coating procedure was repeated, and the pellet was resuspended in acetate buffer saline W pity for final nanocarriers of about lOmg / mL dispersion.

[0巧7] 实例5:含有雷帕霉素的金纳米载体(AuNCs)的制备(预示的) [Qiao 0 7] Example 5: Preparation of Gold vector containing rapamycin (AuNCs) of (Prophetic)

[028引服-PEG-雷帕霉素的制备: [028 serving -PEG- primer rapamycin prepared:

[0289] 将PEG酸二硫化物(1.0当量)、雷帕霉素(2.0-2.5当量)、0〇:(2.5当量)^及01八? [0289] The PEG acid disulfide (1.0 equiv.), Rapamycin (2.0-2.5 equiv), 0〇: (2.5 equiv.) ^ 01 and eight? (3.0当量)在无水DMF中的溶液在室溫下揽拌过夜。 (3.0 equiv.) In dry DMF was stirred at room temperature overnight embrace. 通过过滤去除不溶的二环己脈并且将滤液添加到异丙醇(IPA)中W沉淀出PEG-二硫化物-二-雷帕霉素醋并用IPA洗涂并干燥。 Removed by filtration and the insoluble dicyclohexyl pulse filtrate was added to isopropanol (IPA) in W PEG- precipitated disulfide - bis - acetate and rapamycin washcoated with IPA and dried. 聚合物然后用在DMF中的Ξ (2-簇乙基)麟盐酸盐处理,W便将PEG二硫化物还原成硫醇基PEG雷帕霉素醋化S-PEG-雷帕霉素)。 Then the polymer in DMF Ξ (2- ethyl cluster) hydrochloride and Lin, W put PEG thiol to a disulfide reducing rapamycin PEG esterified rapamycin S-PEG-) . 通过沉淀从IPA回收生成的聚合物并且如先前所述进行干燥并且通过Η NMR和GPC来分析。 And dried as described previously recovered by precipitation of the polymer produced from IPA and by Η NMR and GPC analyzes.

[0290] 金NCs(AuNCs)的形成: [0290] Gold NCs (AuNCs) is formed:

[0291] 在一个装备有冷凝器的1L圆底烧瓶中,在剧烈揽拌下加热回流500mL的ImM HAuC14的水溶液10分钟。 [0291] In a 1L round bottom flask equipped with a condenser, was heated at reflux with vigorous embrace stirred solution of 500mL ImM HAuC14 for 10 minutes. 然后向揽拌的溶液迅速添加50mL的40mM的巧樣酸Ξ钢溶液。 Qiao comp acid was then added a solution of 50mL of 40mM Ξ steel to embrace rapidly stirred solution. 将生成的深酒红色溶液保持回流25-30分钟,然后停止加热,并且冷却该溶液至室溫。 The resulting dark wine red solution was maintained at reflux for 25-30 minutes and then heating was discontinued, and the solution was cooled to room temperature. 然后通过一个0.祉m薄膜过滤器过滤该溶液,W给出AuNCs溶液。 The solution was then filtered through a 0.5 m membrane filter Zhi, W is given AuNCs solution. 使用可见光谱学和透射电子显微术表征AuNCs。 Using visible spectroscopy and transmission electron microscopy to characterize AuNCs. 运些AuNCs直径大约20nm,被巧樣酸盐包裹,并且吸收峰在520nm处。 These transport AuNCs about 20 nm in diameter, is wrapped clever like salt, and an absorption peak at 520nm.

[0巧2] 具有服-PEG-雷帕霉素的AuNCs共辆物: [Qiao 2 0] having a service -PEG- rapamycin AuNCs was co vehicle:

[0293]将150μ1的服-阳G-雷帕霉素的溶液(在lOmM抑9.0碳酸盐缓冲液中的ΙΟμΜ)添加到ImL的20nm直径的巧樣酸盐包裹的金纳米载体(1.16ηΜ)中,W产生2500:1的硫醇基与金的摩尔比。 [0293] The 150μ1 of clothing - the male G- rapamycin solution (inhibition ΙΟμΜ carbonate buffer 9.0 in 10mM) was added to the salt-like wrapped clever 20nm diameter gold carrier ImL of (1.16ηΜ ), W 2500 is generated: the molar ratio of thiol group and gold. 1. 在室溫在氣下揽拌混合物1小时W允许金纳米载体上的硫醇与巧樣酸盐完全交换。 Gas at room temperature under W embrace stirred mixture for 1 hour to allow the carrier and the thiol on the gold salt clever comp complete exchange. 然后通过在12,000g下离屯、30分钟将表面上的具有PEG-雷帕霉素的AuNCs纯化。 Then, 30 minutes AuNCs rapamycin having PEG- purified from the village on the surface by at 12,000g. 将上清液倾析出来并且然后将含有AuNC-S-PEG-雷帕霉素的沉淀用lx PBS缓冲液沉淀洗涂。 The supernatant is decanted off and then AuNC-S-PEG- containing rapamycin precipitate was precipitated with lx PBS wash buffer coating. 然后将纯化的金-PEG-雷帕霉素纳米载体再悬浮于合适的缓冲液中W便进行进一步分析和生物测定。 The purified rapamycin -PEG- gold nano-carriers were resuspended in a suitable buffer for further analysis and W will bioassay.

[0巧4] 实例6:含有卵白蛋白的介孔二氧化娃-金核-壳纳米载体(预示的) [Qiao 0 4] Example 6: baby mesoporous silica containing ovalbumin - gold core - shell nanocarriers (Prophetic)

[OWS]介孔Si〇2纳米颗粒核屯、通过溶胶-凝胶法来产生。 [The OWS] Mesoporous Si〇2 Tun nanoparticle core, by the sol - gel method to produce. 将十六烷基Ξ甲基漠化锭(CTAB) (0.5g)溶解在去离子水(500mL)中,并且然后将2M的化0H水溶液(3.5血)添加到CTAB 溶液中。 The methyl hexadecyl Ξ desertification ingot (CTAB) (0.5g) was dissolved in deionized water (500 mL), and then a 2M aqueous solution of 0H (3.5 of blood) was added to the CTAB solution. 将该溶液揽拌30分钟,并且然后向该溶液中添加四乙氧基硅烷(TE0S) (2.5mL)。 The solution was stirred for 30 minutes embrace, and then tetraethoxysilane (TE0S) (2.5mL) to this solution. 在80°C的溫度下将生成的凝胶揽拌3小时。 At a temperature to 80 ° C and the resulting gel embrace for 3 hours. 通过过滤捕获形成的白色沉淀,随后用去离子水洗涂并且在室溫下干燥。 By capturing the white precipitate formed was filtered, washed with deionized water and then coated and dried at room temperature. 然后通过悬浮于HC1的乙醇溶液中过夜而从颗粒中提取出残留的表面活性剂。 It is then extracted by suspension in a solution of HC1 in ethanol overnight from the particles of the residual surfactant. 将运些颗粒用乙醇洗涂、离屯、、并且在超声处理下再分散。 The transport of these particles are coated washed with ethanol, and then dispersed from the village ,, under sonication. 运个洗涂程序另外再重复两次。 Further transport a wash coating procedure was repeated twice.

[0296] 然后使用(3-氨基丙基)-Ξ乙氧基硅烷(APTMS)用氨基对Si化纳米颗粒进行功能化。 [0296] then (3-aminopropyl) -Ξ ethoxysilane (the APTMS) be functionalized with amino Si nanoparticles. 为此,将颗粒悬浮在乙醇(3〇1^)中,并且将4口了18(5化山添加到悬浮液中。允许该悬浮液在室溫下静置2小时,并且然后煮沸4小时,通过周期性地添加乙醇使体积保持恒定。残留的反应物通过五个离屯、洗涂和再分散于纯乙醇中的循环来去除。 For this purpose, the particles suspended in ethanol (3〇1 ^), and adding 4 of 18 (5 of mountain to the suspension. The suspension was allowed to stand at room temperature for 2 hours and then boiled for 4 hours by adding ethanol periodically to maintain a constant volume. the residue from the reaction was purified by five Tun, wash coating, and redispersed in pure ethanol to remove the loop.

[0297] 在一个单独的反应中,产生l-4nm直径的金种子。 [0297] In a separate reaction, to produce gold seed l 4nm-diameter. 在此反应中使用的所有的水首先被去离子并且然后从玻璃蒸馈。 All water used in this reaction is first distilled and then fed to an ion from the glass. 将水(45.5mL)添加到lOOmL圆底烧瓶中。 Water (45.5 mL) was added to a lOOmL round bottom flask. 在揽拌下,添加0.2M水性化0H( 1.5mL),随后添加氯化四(径甲基)鱗(THPC)的1 %水溶液(1. OmL)。 In embrace mix was added 0.2M aqueous based 0H (1.5mL), followed by the addition tetrachloride (meth diameter) scale (of THPC) 1% aqueous solution (1. OmL). 在添加THPC溶液之后两分钟,添加已经老化至少15分钟的氯金酸的lOmg/mL水溶液(2mL)。 lOmg / mL solution (2mL) two minutes after the addition was added solution THPC has been aged for at least 15 minutes chloroauric acid. 通过用水透析来纯化运些金种子。 These transport SEED purified by dialysis against water.

[0298] 为了形成核-壳纳米载体,将W上形成的氨基功能化的Si化纳米颗粒首先在室溫下与运些金种子混合2小时。 [0298] In order to form a core - shell nanocarriers, amino functionalized nanoparticles formed of Si on the W first at room temperature and transported more gold seeds mixed for 2 hours. 将金修饰的Si化颗粒通过离屯、来收集,并且与氯金酸和碳酸氨钟的水溶液混合来形成金外壳。 Si-modified gold particles by ion Tun, collected, and mixed with an aqueous solution of chloroauric acid and ammonium carbonate to form a gold bell housing. 然后将运些颗粒离屯、洗涂、并且再分散于水中。 These particles are then transported from the village, wash coating, and redispersed in water. 通过将运些颗粒悬浮于硫醇化的卵白蛋白溶液(Img/L)中72小时来负载硫醇化的卵白蛋白(通过用2- 亚氨基硫烧盐酸盐处理卵白蛋白而制成)。 Will be transported by these particles are suspended in a solution of thiolated ovalbumin (Img / L) for 72 hours to load thiolated ovalbumin (by using 2-iminothiolane hydrochloride burning process ovalbumin made). 颗粒然后用lx PBS(pH 7.4)沉淀洗涂W去除游离蛋白。 Particles are then treated with lx PBS (pH 7.4) W precipitate was washed to remove free protein coating. 然后将生成的含有卵白蛋白的二氧化娃-金核-壳纳米载体再悬浮于lx PBS中W便进行进一步分析和测定。 And then the resultant baby dioxide containing ovalbumin - gold core - shell nanocarriers are resuspended in lx PBS W will be further analyzed and measured.

[0巧9] 实例7:含有雷帕霉素和卵白蛋白的脂质体(预示的) [Qiao 0 9] Example 7: containing rapamycin and ovalbumin liposomes (Prophetic)

[0300]运些脂质体通过薄膜水合来形成。 [0300] Some transportation liposomes formed by a thin film hydration. 将1,2-二栋桐酷-sn-甘油基-3憐酸胆碱化PPC) (32皿〇1)、胆固醇(32μπιο 1)、W及雷帕霉素(6.4μπιο1)溶解在纯氯仿(3mL)中。 1,2 Kazuto cool -sn- glycero two acid choline -3 pity of PPC) (32 〇1 dish), cholesterol (32μπιο 1), W, and rapamycin (6.4μπιο1) was dissolved in pure chloroform (3mL) in. 将此脂质溶液添加到lOmL玻璃管中,并且在氮气流下去除溶剂并且真空干燥6小时。 This lipid solution was added to lOmL glass tube, and the solvent was removed under a stream of nitrogen and dried in vacuo for 6 hours. 通过用2.0ml的25mM MOPS缓冲液抑8.5使该膜水合来获得多层囊泡,其含有过量的卵白蛋白。 By buffer 25mM MOPS 2.0ml 8.5 The inhibition membrane hydration to obtain a multilamellar vesicle, which contains an excess of ovalbumin. 使管满旋,直到脂质膜从管表面上剥落。 Full rotation of the tube, until a lipid film is peeled off from the tube surface. 为了将多层囊泡打破成单层的,应用十个冷冻(液氮)和融化(30°C水浴)循环。 In order to break multilamellar vesicles into a single, application ten frozen (liquid nitrogen) and thawing (30 ° C water bath) cycle. 然后将样品在25mM MOPS缓冲液抑8.5中稀释到1ml。 The sample was then buffer 25mM MOPS 8.5 suppressor diluted to 1ml. 通过使样品10倍穿过200皿孔聚碳酸醋过滤器而挤出而将生成的脂质体的大小均匀化。 And the size of the resulting liposomes homogenized by 10 times the sample passes through the aperture 200 dishes extruded polycarbonate filter. 然后使用生成的脂质体用于进一步分析和生物测定。 Liposomes then generated for further analysis and bioassay.

[030。 [030. 实例8:由具有表面共辆的卵白蛋白的修饰聚氨基酸组成的聚合纳米载体(预示的) Example 8: polymerization of a nanocarrier having a surface co-polyamino acid modified vehicle consisting of ovalbumin (Prophetic)

[0302] 步骤1.用心苯丙氨酸乙醋化斗46)修饰的聚(丫-谷氨酸)(丫斗64)的制备:将4.7 单位毫摩尔的丫-PGA (Μη = 300kD)溶解在0.3N-NaHC〇3水溶液(50mL)中。 [0302] Step 1. intentions phenylalanine ethyl ester of bucket 46) modified poly (Ah - glutamic acid) (Ah bucket 64): A mixture of 4.7 mmol of units Ya -PGA (Μη = 300kD) was dissolved in NaHC〇3 0.3N-aqueous solution (50mL). 将kPAE (4.7mmo 1) 和抓C.肥1 (4.7mmol)添加到该溶液中并且在4°C下揽拌30分钟。 The kPAE (4.7mmo 1) and grip fertilizer C. 1 (4.7mmol) was added to the solution and stirred embrace at 4 ° C 30 min. 然后在揽拌下将该溶液保持在室溫24小时。 The solution was then stirred at embrace kept at room temperature for 24 hours. 使用具有MWC0 50kD的透析薄膜通过渗析去除低分子量化学物质。 Having MWC0 50kD dialysis film is removed by dialysis of low molecular weight chemicals. 通过冻干获得生成的丫-PGA-接枝-L-PAE。 Ah -PGA- obtained graft -L-PAE generated by lyophilization.

[0303] 步骤2.由丫丫-PGA-接枝-L-PAE聚合物制备纳米颗粒:由丫-PGA-接枝-L-PAE组成的纳米颗粒通过沉淀和透析方法来制备。 [0303] Step 2. By Ya-Ya -PGA- -L-PAE graft polymer nanoparticles prepared by: Ah -PGA- graft -L-PAE nanoparticle composition prepared by precipitation and dialysis. 将丫-PGA-接枝-kPAE(20mg)溶解在2ml的DMS0 中,随后添加2mL的水W形成半透明溶液。 The graft Ah -PGA- -kPAE (20mg) was dissolved in 2ml of DMS0, followed by addition of water W 2mL of a translucent solution. 然后在室溫下用蒸馈水使用纤维素薄膜配管(50, 000MWC0)透析该溶液72小时,W形成纳米颗粒并且去除有机溶剂。 It is then fed at room temperature with distilled water using a cellulose film pipe (50, 000MWC0) the solution was dialyzed 72 hours, W nanoparticles formed and removing the organic solvent. 蒸馈水W12小时的间隔进行更换。 Distilled water feeding hour intervals W12 replaced. 然后使用生成的纳米颗粒溶液(在水中的lOmg/mUW用于抗原共辆。 Then using the generated nanoparticles solution (lOmg in water / mUW vehicle for co antigen.

[0304] 步骤3.卵白蛋白与丫丫-PGA纳米颗粒的共辆:丫丫-PGA纳米颗粒(lOmg/ml)的表面簇酸基团首先在环境溫度下通过抓C和NHS(各自在憐酸盐缓冲液中的lOmg/mL,抑5.8) 活化2小时。 [0304] Step 3. Ovalbumin nanoparticles and Ya-Ya -PGA total vehicle: Ya-Ya -PGA surface of the cluster acid groups nanoparticles (lOmg / ml) and C is first caught by NHS (each at ambient temperature in a pity formate buffer lOmg / mL, inhibiting 5.8) activated for 2 hours. 在沉淀洗涂W去除过量的EDC/NHS之后,将活化的纳米颗粒与1 mL的卵白蛋白(lOmg/ml)混合在憐酸盐缓冲盐水(PBS,pH 7.4)中,并且将混合物在4°08°(:下解育24小时。将生成的卵白蛋白共辆的丫丫-PGA纳米颗粒用PBS洗涂两次并且W5mg/mL再悬浮于PBS 中W便进行进一步分析和生物测定。 After the precipitate was washed to remove excess coating W EDC / NHS, activated nanoparticles with 1 mL of ovalbumin (lOmg / ml) were mixed in pity-buffered saline (PBS, pH 7.4), and the mixture was 4 ° 08 ° (: the solution for 24 hours to generate a total vehicle ovalbumin -PGA Ya-Ya-coated nanoparticles washed twice with PBS and W5mg / mL were resuspended in PBS for further analysis and W will bioassay.

[03化]实例9:红细胞生成素化P0)封装的丫-PGA纳米颗粒(预示的) [Of 03] Example 9: Erythropoietin of P0) -PGA Ah encapsulated nanoparticles (Prophetic)

[0306] 为了制备EP0封装的丫-PGA纳米颗粒,将0.25-4mg的EP0溶解在1血的PBS(抑7.4) 中,并且将ImL的丫-PGA-接枝-kPAE (DMS0中的1 Omg/mL)添加到EP0溶液中。 [0306] In order -PGA Ah encapsulated nanoparticles prepared EP0, the EP0 0.25-4mg dissolved in the blood of PBS 1 (inhibition 7.4), and the graft ImL Ya -PGA- -kPAE (DMS0 of 1 Omg / mL) was added to the solution EP0. 将生成的溶液在14,000xg下离屯、15分钟并且用PBS重复清洗。 The resulting solution from the village, 15 min and repeatedly washed with PBS at 14,000xg. 然后将生成的EP0封装的丫-PGA纳米颗粒再悬浮于PBS(5mg/mL)中W便进行进一步分析和生物测定。 And then the resultant EP0 -PGA Ah encapsulated nanoparticles were resuspended in PBS (5mg / mL) will be further analyzed in W and bioassay.

[0307] 实例10:含有卵白蛋白的金纳米载体(AuNCs)的制备(预示的) [0307] Example 10: Preparation of gold-containing ovalbumin carrier (AuNCs) of (Prophetic)

[030引步骤1.金NCs(AuNCs)的形成:在一个装备有冷凝器的1L圆底烧瓶中,在剧烈揽拌下加热回流500mL的ImM HAUC14的水溶液10分钟。 [030 Step 1. Primer gold NCs (AuNCs) is formed: in a 1L round bottom flask equipped with a condenser, was heated at reflux with vigorous embrace stirred solution of 500mL ImM HAUC14 for 10 minutes. 然后向揽拌的溶液迅速添加50mL的40mM 的巧樣酸Ξ钢溶液。 Qiao comp acid was then added a solution of 50mL of 40mM Ξ steel to embrace rapidly stirred solution. 将生成的深酒红色溶液保持回流25-30分钟,然后停止加热,并且冷却该溶液至室溫。 The resulting dark wine red solution was maintained at reflux for 25-30 minutes and then heating was discontinued, and the solution was cooled to room temperature. 然后通过一个0.祉m薄膜过滤器过滤该溶液,W给出AuNCs溶液。 The solution was then filtered through a 0.5 m membrane filter Zhi, W is given AuNCs solution. 使用可见光谱学和透射电子显微术表征AuNCs。 Using visible spectroscopy and transmission electron microscopy to characterize AuNCs. 运些AuNCs直径大约20nm,被巧樣酸盐包裹,并且吸收峰在520nm处。 These transport AuNCs about 20 nm in diameter, is wrapped clever like salt, and an absorption peak at 520nm.

[0309]步骤2 .卵白蛋白与AuNCs的共辆:将150μ1的硫醇化的卵白蛋白溶液(在1 OmM pH 9.0碳酸盐缓冲液中,ΙΟμΜ)添加至IjImL的20皿直径的巧樣酸盐包裹的金纳米载体(1.16nM) 中,W产生2500:1的硫醇基与金的摩尔比。 . [0309] Step 2 with ovalbumin AuNCs co vehicle: The thiolated ovalbumin 150μ1 solution (at 1 OmM pH 9.0 carbonate buffer, ΙΟμΜ) was added to 20 IjImL diameter dish-like salt Qiao gold carrier package (1.16nM), W is generated 2500: thiol group molar ratio of 1 gold. 在室溫在氣下揽拌混合物1小时W允许金纳米载体上的硫醇与巧樣酸盐完全交换。 Gas at room temperature under W embrace stirred mixture for 1 hour to allow the carrier and the thiol on the gold salt clever comp complete exchange. 然后通过在12,000g下离屯、30分钟将表面上的具有卵白蛋白的AuNCs纯化。 Then, 30 minutes AuNCs with ovalbumin purified from the village on the surface by at 12,000g. 将上清液倾析出来并且然后将含有AuNC-卵白蛋白的沉淀用lx PBS缓冲液沉淀洗涂。 The supernatant is decanted off and then the precipitate containing AuNC- ovalbumin with lx PBS buffer wash coating precipitate. 然后将纯化的金-卵白蛋白纳米载体再悬浮于合适的缓冲液中W便进行进一步分析和生物测定。 The purified gold - ovalbumin nanocarriers resuspended in a suitable buffer for further analysis and W will bioassay.

[0310] 实例η:通过T细胞表型分析评估致耐受性免疫应答(预示的) [0310] Example η: Assessment Analysis immune response induced tolerance (Prophetic) via the T cell phenotype

[03川将本发明的组合物溶解憐酸盐缓冲盐水(PBS)中并且W含SOOiig的该组合物的0.1-0.2ml肌内地注射到雌路易斯鼠中。 [03 Chuan composition of the present invention is dissolved Rei-buffered saline (PBS) and the composition containing the W SOOiig 0.1-0.2ml of intramuscularly injected into female Lewis rats. 一个大鼠对照组接收0.1-0.2ml的PBS。 A control group of rats received PBS 0.1-0.2ml of. 在注射之后9到10天,从运些小鼠收获脾脏和淋己结并且通过经由40WI1的尼龙细胞滤网浸软组织来获得单细胞悬浮液。 After injection of 9-10 days, and single cell suspensions were obtained from mouse spleens were harvested and transported these lymph nodes via hexyl nylon cell strainer 40WI1 of macerated tissue. 在PBS(1%FCS)中对样品进行染色,并且适当地稀释相关单克隆抗体。 Samples were stained in PBS (1% FCS), and diluted appropriately associated monoclonal antibody. 舰化丙晚染色细胞从分析中排除。 Night propan ship of stained cells were excluded from analysis. 在LSR2流式细胞仪(抓生物科学公司(BD Biosciences), USA)上获得样品并且使用FACS Diva软件进行分析。 In LSR2 flow cytometer (catch Biosciences (BD Biosciences), USA) to obtain the sample using FACS Diva software for analysis. 在细胞上分析标记CD4、CD8等的表达。 Analysis of expression of markers CD4, CD8, etc. on the cell. 例如CD4巧细胞和/或CD8巧细胞的存在降低表明了所希望的免疫应答的诱导。 Qiao such as CD4 cells and / or the presence of CD8 cells is reduced clever show induction of the desired immune response.

[0312] 实例12:体内评估针对自身抗原的致耐受性免疫应答(预示的) [0312] Example 12: in vivo evaluation against self tolerogenic immune response to an antigen (Prophetic)

[0313] 对Ba化/c小鼠给予包含MHC I类限制性表位和/或MHC II类限制性表位的一种自身抗原,并且分别评定CD8巧细胞和/或CD4巧细胞增殖的水平。 [0313] based on Ba / c mice were administered comprises one MHC class I restricted epitope and / or MHC II-restricted epitopes of the autoantigen, and the level of proliferation were assessed clever CD8 cells and / or CD4 cells clever . 随后,W剂量依赖性方式皮下给予本发明的组合物,该组合物包含含有运些表位的该自身抗原或其部分W及一种免疫抑制剂。 Subsequently, W subcutaneously administered dose dependent manner the composition of the invention, the composition comprises the operation some autoantigen epitope or a portion W, and an immunosuppressant. 再次分别评定CD8巧细胞和/或CD4巧细胞增殖的水平,其中增殖降低表明一种致耐受性免疫应答。 Evaluation of CD8 cells were again clever and / or the level of CD4 cell proliferation Coincidentally, wherein one kind show decreased proliferation tolerogenic immune response.

[0314] 实例13:体内降低针对可移植的骨髓细胞的所不希望的免疫应答(预示的) [0314] Example 13: in vivo to reduce an immune response against the transplanted bone marrow cells may be undesirable (Prophetic)

[0315] 在受试者接受骨髓移植前四周,向该受试者皮下给予一个群体的至少106个合成纳米载体,运些合成纳米载体包含免疫抑制剂和从骨髓细胞中获得或衍生出的多种抗原。 [0315] around a group subcutaneously administered to the subject before the subject received a bone marrow transplant at least 106 synthetic nanocarriers, shipped some synthetic nanocarrier comprising an immunosuppressant and obtained or derived from bone marrow cells of a multi- antigens. 在该受试者接受该移植之后,在接受该移植后的第一周期间内每日一次、并且然后在接下来Ξ周每周一次、并且然后在接下来11周每个月一次地对该受试者中的所不希望的免疫应答的产生进行评定。 After the subjects receiving the graft, once during the first week after receiving the transplant day, and then once a week for the next Ξ weeks, and then once a month for the next 11 weeks generating subject undesired immune responses were evaluated. 作为评定的一部分,取得CD8巧细胞和/或CD4巧细胞计数,并且与向该受试者给予骨髓移植或运些合成纳米载体之前取得的CD8巧细胞和/或CD4巧细胞计数进行比较。 As part of assessment made clever CD8 cells and / or CD4 cell count Qiao, Qiao CD8 cells obtained prior to transplantation or transported and some synthetic nanocarriers administering to the subject the bone marrow and / or CD4 cell count compared skillfully. 在第一年期间,向该受试者双月地给予维持剂量的运些合成纳米载体。 During the first year, bimonthly to the subject administered a maintenance dose of these synthetic nanocarriers operation. 预期该受试者不展现或仅展示极少量对骨髓移植特异的CD8巧细胞和/或CD4巧细胞水平。 The subject is expected to exhibit no or only very small amounts of impressions and / or the level of CD4 cells Qiao transplanted bone marrow cells specific CD8 clever.

[0引引实例14:评估效应性T细胞的体外缺失(预示的) [Cited Cited 0 Example 14: In vitro assessment of effector T cells in absence (Prophetic)

[0317]可W使包含效应性T细胞或效应性T细胞前体的一个细胞群体在体外与一种在此提供的组合物接触。 [0317] W can make a cell population comprising effector T cell precursor effector T cells, or contacting in vitro a composition provided herein with one. 在足W使该组合物与该细胞群体中的运些效应性T细胞或效应性T细胞前体相互作用的一段时间后,预期效应性T细胞数目的降低。 After a sufficient period of time W so that the composition with the cell population before these effector T cells or shipped effector T cell interactions, and the desired number of effector T cells is reduced. 在一些实施例中,足W使该群体中的效应性T细胞数目降低的一段时间是约1天、约2天、约3天、约4天、约5天、约6天、约1 周、约2周、约3周、或约4周。 In some embodiments, the foot W so reducing the number of effector T cells in the population of a period of time is about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 1 week , about 2 weeks, about 3 weeks, or about 4 weeks. 在一些实施例中,在该时间段后效应性T细胞的数目降低,例如降低到该群体中效应性T细胞的原始总数目或相对数目的小于约50%、小于约60%、小于约70%、小于约80%、小于约90%、小于约95 %、小于约98 %、或小于约99 %。 In some embodiments, the number of effector T cells is reduced after this period of time, for example to less than about 50% of the original total number or relative number of this population of effector T cells, less than about 60%, less than about 70 %, less than about 80%, less than about 90%, less than about 95%, less than about 98%, or less than about 99%. 在一些实施例中, 在该时间段后效应性T细胞完全从该细胞群体中消失。 In some embodiments, the effector T cells disappear from the cell population after the time period. 在一些实施例中,在使该细胞群体与在此提供的一种组合物接触之前测定该群体中效应性T细胞的总数目和/或相对数目,W便建立该群体中效应性T细胞的基线数目。 In some embodiments, contacting the cell population in contact with a composition provided herein is prior to the total number and / or the relative number of effector T cell population was determined, W is established in this population of effector T cells The number of baseline. 在一些实施例中,使该细胞群体与在此提供的一种组合物接触一次。 In some embodiments, the cell population so that the contacting time with a composition provided herein. 在一些实施例中,使该细胞群体与在此提供的一种组合物重复接触。 In some embodiments, the cell population so that the repeated contact with a composition provided herein.

[031引还在接触之后确定该细胞群体中效应性T细胞的数目和/或存在。 [031 primer is also determined after contacting the cell population number of effector T cells and / or presence. 在一些实施例中,监测一段时间内效应性τ细胞的数目和/或存在,例如通过执行多个随后的效应性τ细胞检测测定。 In some embodiments, the number of the monitoring period τ effector cells and / or the presence of, for example, a plurality of subsequent detection τ determined by performing effector cells. 在一些实施例中,通过从该细胞群体取得代表该细胞群体的一个样品、用特异性地检测效应性Τ细胞标记的抗体或染色剂对该样品中所含的细胞染色、并且例如通过FACS 或通过免疫组织化学法检测表达该样品中效应性T细胞标记的细胞,来确定该细胞群体中效应性T细胞的数目和/或存在。 In some embodiments, by taking a representative sample of the population of cells from the cell population, cells were stained with specific labeled detecting antibodies or effector cells Τ the stain contained in the sample, for example by FACS or It was detected by the expression of effector T cells in a sample labeled cells by immunohistochemistry to determine the number of the cell population of effector T cells and / or presence. 在一些实施例中,将效应性T细胞定量。 In some embodiments, the effector T cells quantified. 在一些实施例中,将接触之后确定的效应性T细胞的量与接触之前的效应性T细胞的量相比较,例如与效应性T 细胞的基线数目相比较,其中,如果该细胞群体中效应性T细胞的数目在接触后比基线数目更低,则确定已经发生了针对该组合物的一种致耐受性应答。 In some embodiments, the amount of the previous amount determined after contacting the effector T cells in contact with effector T cells as compared to, for example, compared with the baseline number of effector T cells, wherein, if the cell population Effect the number of T cells after exposure to a lower number than the baseline, it is determined to have occurred is directed to a tolerogenic response induced by the composition.

[0319] 实例15:评估效应性T细胞的体内缺失(预示的) [0319] Example 15: In vivo evaluation deletion effector T cells (Prophetic)

[0320] 可W使包含效应性T细胞或效应性T细胞前体的一个细胞群体在体内与一种在此提供的组合物接触。 [0320] W can make a cell population comprising effector T cell precursor effector T cells or in contact with one body composition provided herein. 在一些实施例中,将该组合物或细胞群体给予拥有包含效应性T细胞或效应性T细胞前体的细胞群的受试者。 In some embodiments, the composition has a subject or population of cells comprising administering a population of cells or T cell precursors effect effector T cells. 在足W使该组合物与该受试者中的运些效应性T细胞或效应性T细胞前体相互作用的一段时间后,预期效应性T细胞数目的降低。 After a period of time sufficient to make the composition W interacts with these transportation effector T cells or T cell precursors effector of the subject, the desired number of effector T cells is reduced. 在一些实施例中,足W使该细胞群体中的效应性T细胞数目降低的一段时间是约1天、约2天、约3天、约4 天、约5天、约6天、约1周、约2周、约3周、或约4周。 In some embodiments, the foot W so that a period of time to reduce effector T cell numbers of the cell population is about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 1 weeks, about 2 weeks, about 3 weeks, or about 4 weeks. 在一些实施例中,足W实现效应性T细胞数目降低的该时间段长于4周。 In some embodiments, a sufficient number of effector T W achieve a reduction of cells in the time period longer than four weeks. 在一些实施例中,效应性T细胞的数目在该时间段后降低,例如降低到该受试者中效应性T细胞的原始总数目或相对数目的小于约50%、小于约60%、小于约70 %、小于约80 %、小于约90 %、小于约95 %、小于约98 %、或小于约99 %。 In some embodiments, the number of effector T cells after this period lowered, for example to less than about 50% of the original total number or relative number of the subject of effector T cells, less than about 60%, less than about 70%, less than about 80%, less than about 90%, less than about 95%, less than about 98%, or less than about 99%. 在一些实施例中,在该时间段后,效应性T细胞或效应性T细胞的一个特定群体(例如,祀向一种特定抗原的效应性T细胞)完全从受试者中消失。 In some embodiments, after this period, the effector T cells or a specific population of effector T cells (e.g., to a particular antigen worship effector T cells) from the subject completely disappeared. 在一些实施例中,在对该受试者给予在此提供的一种组合物之前确定该受试者中效应性T细胞的总数目和/或相对数目,W便建立该受试者中效应性T细胞的一个基线数目。 In some embodiments, the total number and / or the relative number of effector T cells in the subject is determined prior to administering to the subject a composition provided herein, W is established in the subject effect a baseline number of cells of T. 在一些实施例中,在此提供的一种组合物被给予受试者一次。 In some embodiments, a composition provided herein is administered to the subject once. 在一些实施例中,在此提供的一种组合物被给予受试者多次,例如,直到在该受试者中观察到所希望的效应性T细胞减少。 In some embodiments, a composition provided herein is administered to a subject multiple times, e.g., until the desired reduction observed effector T cells in the subject.

[0321] 还在给予之后确定该受试者中效应性T细胞的数目和/或存在。 [0321] is also determined in the subject after administration of effector T cell numbers and / or presence. 在一些实施例中, 监测一段时间内效应性T细胞的数目和/或存在,例如通过执行多个随后的效应性T细胞检测测定。 In some embodiments, monitoring a period of time the number of effector T cells and / or the presence of, for example, by performing a plurality of subsequent detection assay effector T cells. 在一些实施例中,通过从该受试者取得代表该受试者中的一个T细胞群体的一个样品(例如外周血样品或淋己样品)、用特异性地检测效应性T细胞标记的抗体或染色剂对该样品中所含的细胞染色、并且例如通过FACS或通过免疫组织化学法检测表达该样品中效应性T细胞标记的细胞,来确定该受试者中效应性T细胞的数目和/或存在。 In some embodiments, a sample by a T cell population in the subject (e.g., peripheral blood sample or a sample leaching hexyl) acquired from the representative of the subject, with specific labeled detecting antibody effector T cells stain or staining the cells contained in the sample, and for example, the expression of T cell marker in a sample of effector cells detected by FACS chemically or by immunohistochemistry to determine the number of the subject of effector T cells, and / or presence. 在一些实施例中, 将效应性T细胞定量。 In some embodiments, the effector T cells quantified. 在一些实施例中,将给予之后确定的效应性T细胞的量与给予之前的效应性T细胞的量相比较,例如与效应性T细胞的基线数目相比较,其中,如果该受试者中效应性T细胞的数目在给予后比基线数目更低,则确定已经发生了针对该组合物的一种致耐受性应答。 The amount of effector T cells, in some embodiments of the previous embodiment, the amount determined after administration of effector T cells compared with the administration of, for example, compared with the baseline number of effector T cells, wherein, if the subject the number of effector T cells after administration of a lower number than the baseline, it is determined to have occurred is directed to a tolerogenic response induced by the composition.

[0巡]实例16:体内评估使用包含免疫抑制剂和APC可呈递抗原的合成纳米载体的致耐受性免疫应答 Tolerance induced immune responses in vivo was assessed using synthetic nanocarriers comprising an immunosuppressant and may present antigen APC: [0 patrol] Example 16

[0323] 合成纳米载体生产的材料和方法 [0323] Materials and Methods Production of synthetic nanocarriers

[0324] 纳米载体1 [0324] 1 nanocarrier

[0325] 从TSZ化学品公司(威尔逊街185号,弗雷明汉,马萨诸塞州01702;产品目录# R1017)购买雷帕霉素。 [0325] from the TSZ Chemicals (185 Wilson Street, Framingham, MA 01702; catalog # R1017) purchase of rapamycin. 从SurModics制药公司(汤姆马下路756号,伯明翰,亚拉己马州35211;产品代码7525化G 7A)购买丙交醋:乙交醋比为3:1并且特性粘度为0.75化/g的化GA。 Pharmaceuticals from SurModics (Tang Muma drop 756, Birmingham, hexyl Maryland 35211; product code 7525 of G 7A) for later vinegar lactide: glycolide ratio of acetate 3: 1 and an intrinsic viscosity of 0.75 / g, of GA. 合成了阳G嵌段大约为5,OOODa并且PLA嵌段大约为20,000化的PLA-PEG嵌段共聚物。 Yang G blocks were synthesized about 5, OOODa and the PLA block of about 20,000 PLA-PEG block copolymer. 从EMD化学品公司(产品编号1.41350.1001)购买聚乙締醇(85 % -89 %水解的)。 Polyvinyl alcohol for later association from the EMD Chemicals (Catalog No .: 1.41350.1001) (85% -89% hydrolyzed).

[0326] 如下制备溶液: [0326] A solution was prepared as follows:

[0327] 溶液1:在二氯甲烧中的雷帕霉素@50mg/mL。 [0327] Solution 1: A burning in methylene rapamycin @ 50mg / mL. 通过将雷帕霉素溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by dissolving the pure rapamycin dichloromethane calcining.

[032引溶液2:在二氯甲烧中的PLGA@100mg/mL。 [032 Primer solution 2: PLGA @ 100mg of burning in dichloromethane / mL. 通过将PLGA溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by dissolving the PLGA in neat dichloromethane calcining.

[03巧]溶液3:在二氯甲烧中的PLA-PEG@100mg/mL。 [Qiao 03] Solution 3: dichloromethane burning in the PLA-PEG @ 100mg / mL. 通过将化A-PEG溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by of A-PEG was dissolved in dichloromethane pure calcining.

[0330] 溶液4:在lOOmM的抑8的憐酸盐缓冲液中的聚乙締醇@50mg/mL。 [0330] Solution 4: pity in acetate buffer lOOmM t08 in the association of alcohol polyethylene @ 50mg / mL.

[0331 ]使用水包油乳液来制备纳米载体。 [0331] was prepared using the nanocarriers oil in water emulsion. 通过将溶液1 (0.2mL)、溶液2(0.75mL)、溶液3 (0.25mL)、W及溶液4(3mL)合并在一个小的压力管中并且使用布兰森数字超声波仪250在30%振幅下进行声处理60秒来制备0/W乳液。 By the solution 1 (0.2mL), a solution of 2 (0.75mL), a solution of 3 (0.25mL), W and a solution of 4 (3mL) were combined in a small pressure tube and using Branson Digital Sonifier 250 at 30% the amplitude sonicated for 60 seconds prepared 0 / W emulsion. 将该0/W乳液添加到含有70mM的抑如溝酸盐缓冲溶液(30mL)的烧杯中并且在室溫下揽拌2小时W允许二氯甲烧蒸发并且允许纳米载体形成。 The 0 / W emulsion was added to a beaker containing 70mM suppressing grooves as formate buffer solution (30mL) of W and embrace for 2 hours at room temperature to allow evaporation of dichloromethane and allowed to burn nanocarriers are formed. 通过W下方式来洗涂一部分纳米载体:将纳米载体悬浮液转移至一个离屯、管中,并且在21,000Xg和4°C下离屯、45分钟、去除上清液、并且将沉淀再悬浮于憐酸盐缓冲盐水中。 By way of a portion of the W-coated to wash the nanocarrier: The nanocarrier suspension was transferred to a village from the tube, and 21,000Xg at 4 ° C from the village and 45 minutes, the supernatant was removed, and the pellet was pity salt was suspended in buffered saline. 重复该洗涂程序,并且将沉淀再悬浮于憐酸盐缓冲盐水中W用于约lOmg/mL的最终的纳米载体分散液。 The wash-coating procedure was repeated, and the pellet was resuspended in acetate buffer saline W pity for final nanocarriers of about lOmg / mL dispersion.

[0332] 通过动态光散射来确定纳米载体大小。 [0332] to determine the size of the nanocarrier by dynamic light scattering. 通过HPLC分析来确定在纳米载体中的雷帕霉素的量。 To determine the amount of rapamycin in the nanocarrier by HPLC analysis. 通过重量法来确定每mL悬浮液的总的干燥纳米载体质量。 Determining a total dry mass per mL nanocarrier suspension by the gravimetric method.

[0333] [0333]

Figure CN105999295AD00601

[0334] 纳米载体2 [0334] 2 nanocarrier

[0335] 从己亨美洲公司(柏市街3132号,托伦斯,加利福尼亚州90505,区号#4065609)购买卵白蛋白肤323-339,即一种已知为卵白蛋白的T和B细胞表位的17氨基酸肤。 [0335] purchase ovalbumin own skin 323-339 from Hang Americas (No. 3132 Kashiwa Street, Torrance, California 90505, area code # 4065609), that is known as the ovalbumin T and B cell epitopes 17 amino acids skin. 从SurModics制药公司(汤姆马下路756号,伯明翰,亚拉己马州35211;产品代码7525化G 7A) 购买丙交醋:乙交醋比为3:1并且特性粘度为0.75化/g的化GA。 Pharmaceuticals from SurModics (Tang Muma drop 756, Birmingham, hexyl Maryland 35211; product code 7525 of G 7A) for later vinegar lactide: glycolide ratio of acetate 3: 1 and an intrinsic viscosity of 0.75 / g, of GA. 合成了PEG嵌段大约为5, OOODa并且化A嵌段大约为20,000Da的化A-PEG嵌段共聚物。 Synthesis of the PEG blocks is about 5, OOODa and A blocks of A-PEG of about 20,000Da of the block copolymer. 从EMD化学品公司(产品编号1.41350.1001)购买聚乙締醇(85%-89%水解的)。 Polyvinyl alcohol for later association from the EMD Chemicals (Catalog No .: 1.41350.1001) (85% -89% hydrolyzed).

[0336] 如下制备溶液: [0336] A solution was prepared as follows:

[0337] 溶液1:在稀释的盐酸水溶液中的卵白蛋白肤323-339@20mg/mL。 [0337] solution 1: aqueous solution of hydrochloric acid diluted skin ovalbumin 323-339 @ 20mg / mL. 通过在室溫下将卵白蛋白肤溶解在0.13M盐酸溶液中而制备该溶液。 The solution prepared in 0.13M hydrochloric acid solution by the skin of ovalbumin dissolved at room temperature.

[033引溶液2:在二氯甲烧中的PLGA@100mg/mL。 [033 Primer solution 2: PLGA @ 100mg of burning in dichloromethane / mL. 通过将PLGA溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by dissolving the PLGA in neat dichloromethane calcining.

[0339] 溶液3:在二氯甲烧中的PLA-PEG@100mg/mL。 [0339] Solution 3: dichloromethane burning in the PLA-PEG @ 100mg / mL. 通过将化A-PEG溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by of A-PEG was dissolved in dichloromethane pure calcining.

[0340] 溶液4:在lOOmM的抑8的憐酸盐缓冲液中的聚乙締醇@50mg/mL。 [0340] Solution 4: pity in acetate buffer lOOmM t08 in the association of alcohol polyethylene @ 50mg / mL.

[Ο%1 ]首先制备第一油包水乳液。 [Ο% 1] First, a first oil emulsion was prepared. 通过将溶液1 (0.2mL)、溶液2 (0.75mL)、w及溶液3 (0.25mL)合并在一个小的压力管中并且使用布兰森数字超声波仪250在50%振幅下进行声处理40秒来制备W1/01。 By the solution 1 (0.2mL), a solution of 2 (0.75mL), w, and a solution (0.25 mL of) 3 were combined in a small pressure tube and using Branson Digital Sonifier 250 at 50% amplitude sonication 40 seconds to prepare W1 / 01. 然后通过将溶液4(3.0mL)与第一W1/01乳液合并、满旋10秒、并且使用布兰森数字超声波仪250在30%振幅下进行声处理60秒来制备第二乳液(W1/01/W2)。 The solution was then passed through 4 (3.0mL) and the first W1 / 01 emulsion were combined, spin full 10 seconds, and using the Branson Digital Sonifier 250 acoustically at 30% amplitude for 60 seconds to prepare a second emulsion (W1 / 01 / W2).

[0342] 将该W1/01/W2乳液添加到含有70mM的pH如溝酸盐缓冲溶液(30mL)的烧杯中并且在室溫下揽拌2小时W允许二氯甲烧蒸发并且允许纳米载体形成。 [0342] The added W1 / 01 / W2 emulsion to the beaker containing the groove as pH 70mM acetate buffer solution (30mL) of W and embrace for 2 hours at room temperature, allowing to burn methylene chloride was evaporated and allowed to form the nanocarrier . 通过W下方式来洗涂一部分纳米载体:将纳米载体悬浮液转移至一个离屯、管中,并且W75,600Xg和4°C离屯、35分钟、去除上清液、并且将沉淀再悬浮于憐酸盐缓冲盐水中。 By way of a portion of the W-coated to wash the nanocarrier: The nanocarrier suspension was transferred to a village from the tube, and W75,600Xg and 4 ° C from the village, 35 minutes, the supernatant was removed, and the pellet was resuspended in Rei-buffered saline. 重复该洗涂程序,并且将沉淀再悬浮于憐酸盐缓冲盐水中W用于约lOmg/mL的最终的纳米载体分散液。 The wash-coating procedure was repeated, and the pellet was resuspended in acetate buffer saline W pity for final nanocarriers of about lOmg / mL dispersion.

[0343] 通过动态光散射来确定纳米载体大小。 [0343] to determine the size of the nanocarrier by dynamic light scattering. 通过HPLC分析来确定在纳米载体中的肤的量。 To determine the amount of skin in the nanocarrier by HPLC analysis. 通过重量法来确定每mL悬浮液的总的干燥纳米载体质量。 Determining a total dry mass per mL nanocarrier suspension by the gravimetric method.

[0344] [0344]

Figure CN105999295AD00611

[0345] 纳米载体3 [0345] 3 nanocarrier

[0%6] 从LKT实验设备公司(大学路西2233号,圣保罗,明尼苏达州55114;产品目录# S3449)购买辛伐他汀。 [0% 6] from LKT laboratory equipment company (2233 West University Avenue, St. Paul, Minnesota 55114; Catalog # S3449) buy simvastatin. 从SurModics制药公司(汤姆马下路756号,伯明翰,亚拉己马州35211;产品代码7525化G 7A)购买丙交醋:乙交醋比为3:1并且特性粘度为0.75化/g的化GA。 Pharmaceuticals from SurModics (Tang Muma drop 756, Birmingham, hexyl Maryland 35211; product code 7525 of G 7A) for later vinegar lactide: glycolide ratio of acetate 3: 1 and an intrinsic viscosity of 0.75 / g, of GA. 合成了阳G嵌段大约为5,OOODa并且PLA嵌段大约为20,000化的PLA-PEG嵌段共聚物。 Yang G blocks were synthesized about 5, OOODa and the PLA block of about 20,000 PLA-PEG block copolymer. 从EMD化学品公司(产品编号1.41350.1001)购买聚乙締醇(85 % -89 %水解的)。 Polyvinyl alcohol for later association from the EMD Chemicals (Catalog No .: 1.41350.1001) (85% -89% hydrolyzed).

[0347]如下制备溶液: [0347] A solution was prepared as follows:

[0;34引溶液1:在二氯甲烧中的辛伐他汀@50mg/mL。 [0; 34 primer solution 1: Burning in dichloromethane simvastatin @ 50mg / mL. 通过将辛伐他汀溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by dissolving pure simvastatin dichloromethane calcining.

[0349] 溶液2:在二氯甲烧中的PLGA@1 OOmg/mL。 [0349] Solution 2: In the burning of dichloromethane PLGA @ 1 OOmg / mL. 通过将PLGA溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by dissolving the PLGA in neat dichloromethane calcining.

[03加]溶液3:在二氯甲烧中的PLA-PEG@100mg/mL。 [Plus 03] Solution 3: dichloromethane burning in the PLA-PEG @ 100mg / mL. 通过将化A-PEG溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by of A-PEG was dissolved in dichloromethane pure calcining.

[0351] 溶液4:在lOOmM的抑8的憐酸盐缓冲液中的聚乙締醇@50mg/mL。 [0351] Solution 4: pity in acetate buffer lOOmM t08 in the association of alcohol polyethylene @ 50mg / mL.

[0352] 使用水包油乳液来制备纳米载体。 [0352] was prepared using the nanocarriers oil in water emulsion. 通过将溶液1 (0.15mL)、溶液2(0.75mL)、溶液3 (0.25mL)、W及溶液4(3mL)合并在一个小的压力管中并且使用布兰森数字超声波仪250在30%振幅下进行声处理60秒来制备0/W乳液。 By the solution 1 (0.15mL), a solution of 2 (0.75mL), a solution of 3 (0.25mL), W and a solution of 4 (3mL) were combined in a small pressure tube and using Branson Digital Sonifier 250 at 30% the amplitude sonicated for 60 seconds prepared 0 / W emulsion. 将该0/W乳液添加到含有70mM的抑如溝酸盐缓冲溶液(30mL)的烧杯中并且在室溫下揽拌2小时W允许二氯甲烧蒸发并且允许纳米载体形成。 The 0 / W emulsion was added to a beaker containing 70mM suppressing grooves as formate buffer solution (30mL) of W and embrace for 2 hours at room temperature to allow evaporation of dichloromethane and allowed to burn nanocarriers are formed. 通过W下方式来洗涂一部分纳米载体:将纳米载体悬浮液转移至一个离屯、管中,并且在75,600 Xg和4°C下离屯、35分钟、去除上清液、并且将沉淀再悬浮于憐酸盐缓冲盐水中。 By way of a portion of the W-coated to wash the nanocarrier: The nanocarrier suspension was transferred to a village from the tube, and at 75,600 Xg and 4 ° C from the village, 35 minutes, the supernatant was removed, and the pellet was resuspended pity salt in buffered saline. 重复该洗涂程序,并且将沉淀再悬浮于憐酸盐缓冲盐水中W用于约lOmg/mL的最终的纳米载体分散液。 The wash-coating procedure was repeated, and the pellet was resuspended in acetate buffer saline W pity for final nanocarriers of about lOmg / mL dispersion.

[0353] 通过动态光散射来确定纳米载体大小。 [0353] to determine the size of the nanocarrier by dynamic light scattering. 通过HPLC分析来确定在纳米载体中的辛伐他汀的量。 To determine the amount of simvastatin in the nanocarrier by HPLC analysis. 通过重量法来确定每mL悬浮液的总的干燥纳米载体质量。 Determining a total dry mass per mL nanocarrier suspension by the gravimetric method.

[0354] [0354]

Figure CN105999295AD00621

[0巧日]纳米载体4 [Qiao Day 0] nanocarrier 4

[0356] 从己亨美洲公司(柏市街3132号,托伦斯,加利福尼亚州90505,区号#4065609)购买卵白蛋白肤323-339,即一种已知为卵白蛋白的T和B细胞表位的17氨基酸肤。 [0356] purchase ovalbumin own skin 323-339 from Hang Americas (No. 3132 Kashiwa Street, Torrance, California 90505, area code # 4065609), that is known as the ovalbumin T and B cell epitopes 17 amino acids skin. 从化学品公司(威尔逊街185号,弗雷明汉,马萨诸塞州01702;产品目录#31017)购买雷帕霉素。 Buy rapamycin; from the chemical company (Catalog # 31017 185 Wilson Street, Framingham, MA 01702). 从SurModics制药公司(汤姆马下路756号,伯明翰,亚拉己马州35211;产品代码7525化G 7Α) 购买丙交醋:乙交醋比为3:1并且特性粘度为0.75化/g的化GA。 Pharmaceuticals from SurModics (Tang Muma drop 756, Birmingham, hexyl Maryland 35211; product code 7525 of G 7Α) vinegar later lactide: glycolide ratio of acetate 3: 1 and an intrinsic viscosity of 0.75 / g, of GA. 合成了PEG嵌段大约为5, OOODa并且化A嵌段大约为20,000Da的化A-PEG嵌段共聚物。 Synthesis of the PEG blocks is about 5, OOODa and A blocks of A-PEG of about 20,000Da of the block copolymer. 从EMD化学品公司(产品编号1.41350.1001)购买聚乙締醇(85%-89%水解的)。 Polyvinyl alcohol for later association from the EMD Chemicals (Catalog No .: 1.41350.1001) (85% -89% hydrolyzed).

[0357] 如下制备溶液: [0357] A solution was prepared as follows:

[0358] 溶液1:在稀释的盐酸水溶液中的卵白蛋白肤323-339@20mg/mL。 [0358] solution 1: aqueous solution of hydrochloric acid diluted skin ovalbumin 323-339 @ 20mg / mL. 通过在室溫下将卵白蛋白肤溶解在0.13M盐酸溶液中而制备该溶液。 The solution prepared in 0.13M hydrochloric acid solution by the skin of ovalbumin dissolved at room temperature.

[0359] 溶液2:在二氯甲烧中的雷帕霉素@50mg/mL。 [0359] Solution 2: Burning in dichloromethane rapamycin @ 50mg / mL. 通过将雷帕霉素溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by dissolving the pure rapamycin dichloromethane calcining.

[0360] 溶液3:在二氯甲烧中的PLGA@100mg/mL。 [0360] solution 3: PLGA @ 100mg of burning in dichloromethane / mL. 通过将PLGA溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by dissolving the PLGA in neat dichloromethane calcining.

[0361] 溶液4:在二氯甲烧中的PLA-PEG@100mg/mL。 [0361] Solution 4: the burning of dichloromethane PLA-PEG @ 100mg / mL. 通过将化A-PEG溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by of A-PEG was dissolved in dichloromethane pure calcining.

[0362] 溶液5:在lOOmM的pH 8的憐酸盐缓冲液中的聚乙締醇@50mg/mL。 [0362] Solution 5: pity in acetate buffer pH lOOmM 8 in association polyvinyl alcohol @ 50mg / mL.

[0363] 首先制备第一油包水乳液。 [0363] First preparing a first water in oil emulsions. 通过将溶液l(0.2mL)、溶液2(0.2mL)、溶液3(0.75mL)、 W及溶液4(0.25mL)合并在一个小的压力管中并且使用布兰森数字超声波仪250在50%振幅下进行声处理40秒来制备W1/01。 By the solution l (0.2mL), was combined 2 (0.2mL), a solution of 3 (0.75mL), W and a solution of 4 (0.25mL) in a small pressure tube and using Branson Digital Sonifier 250 at 50 sonicated% amplitude for 40 seconds, and is prepared W1 / 01. 然后通过将溶液5(3.0mL)与第一W1/01乳液合并、满旋10秒、并且使用布兰森数字超声波仪250在30%振幅下进行声处理60秒来制备第二乳液(W1/01/W2)。 Then the solution was 5 (3.0mL) and the first W1 / 01 emulsion were combined, spin full 10 seconds, and using the Branson Digital Sonifier 250 at 30% amplitude for 60 seconds sonication prepared second emulsion (W1 / 01 / W2).

[0364] 将该W1/01/W2乳液添加到含有70mM的pH如溝酸盐缓冲溶液(30mL)的烧杯中并且在室溫下揽拌2小时W允许二氯甲烧蒸发并且允许纳米载体形成。 [0364] The added W1 / 01 / W2 emulsion to the beaker containing the groove as pH 70mM acetate buffer solution (30mL) of W and embrace for 2 hours at room temperature, allowing to burn methylene chloride was evaporated and allowed to form the nanocarrier . 通过W下方式来洗涂一部分纳米载体:将纳米载体悬浮液转移至一个离屯、管中,并且W21,000Xg和4°C离屯、45分钟、去除上清液、并且将沉淀再悬浮于憐酸盐缓冲盐水中。 By way of a portion of the W-coated to wash the nanocarrier: The nanocarrier suspension was transferred to a village from the tube, and W21,000Xg and 4 ° C from the village, 45 minutes, the supernatant was removed, and the pellet was resuspended in Rei-buffered saline. 重复该洗涂程序,并且将沉淀再悬浮于憐酸盐缓冲盐水中W用于约lOmg/mL的最终的纳米载体分散液。 The wash-coating procedure was repeated, and the pellet was resuspended in acetate buffer saline W pity for final nanocarriers of about lOmg / mL dispersion.

[0365] 通过动态光散射来确定纳米载体大小。 [0365] to determine the size of the nanocarrier by dynamic light scattering. 通过HPLC分析来确定在纳米载体中的肤和雷帕霉素的量。 Skin and to determine the amount of rapamycin in the nanocarrier by HPLC analysis. 通过重量法来确定每mL悬浮液的总的干燥纳米载体质量。 Determining a total dry mass per mL nanocarrier suspension by the gravimetric method.

[0366] [0366]

Figure CN105999295AD00631

[0367] 纳米载体5 [0367] 5 nanocarrier

[0368] 从己亨美洲公司(柏市街3132号,托伦斯,加利福尼亚州90505,区号#4065609)购买卵白蛋白肤323-339,即一种已知为卵白蛋白的T和B细胞表位的17氨基酸肤。 [0368] purchase ovalbumin own skin 323-339 from Hang Americas (No. 3132 Kashiwa Street, Torrance, California 90505, area code # 4065609), that is known as the ovalbumin T and B cell epitopes 17 amino acids skin. 从LKT实验设备公司(大学路西2233号,圣保罗,明尼苏达州55114;产品目录#53449)购买辛伐他汀。 Buy simvastatin; from LKT laboratory equipment company (Catalog # 53449 2233 West University Avenue, St. Paul, Minn. 55114). 从SurModics制药公司(汤姆马下路756号,伯明翰,亚拉己马州35211;产品代码7525化G 7A) 购买丙交醋:乙交醋比为3:1并且特性粘度为0.75化/g的化GA。 Pharmaceuticals from SurModics (Tang Muma drop 756, Birmingham, hexyl Maryland 35211; product code 7525 of G 7A) for later vinegar lactide: glycolide ratio of acetate 3: 1 and an intrinsic viscosity of 0.75 / g, of GA. 合成了PEG嵌段大约为5, OOODa并且化A嵌段大约为20,000Da的化A-PEG嵌段共聚物。 Synthesis of the PEG blocks is about 5, OOODa and A blocks of A-PEG of about 20,000Da of the block copolymer. 从EMD化学品公司(产品编号1.41350.1001)购买聚乙締醇(85%-89%水解的)。 Polyvinyl alcohol for later association from the EMD Chemicals (Catalog No .: 1.41350.1001) (85% -89% hydrolyzed).

[0369] 如下制备溶液: [0369] A solution was prepared as follows:

[0370] 溶液1:在稀释的盐酸水溶液中的卵白蛋白肤323-339@20mg/mL。 [0370] solution 1: aqueous solution of hydrochloric acid diluted skin ovalbumin 323-339 @ 20mg / mL. 通过在室溫下将卵白蛋白肤溶解在0.13M盐酸溶液中而制备该溶液。 The solution prepared in 0.13M hydrochloric acid solution by the skin of ovalbumin dissolved at room temperature.

[0371] 溶液2:在二氯甲烧中的辛伐他汀@50mg/mL。 [0371] Solution 2: Burning in dichloromethane simvastatin @ 50mg / mL. 通过将辛伐他汀溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by dissolving pure simvastatin dichloromethane calcining.

[0372] 溶液3:在二氯甲烧中的PLGA@100mg/mL。 [0372] solution 3: PLGA @ 100mg of burning in dichloromethane / mL. 通过将PLGA溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by dissolving the PLGA in neat dichloromethane calcining.

[0373] 溶液4:在二氯甲烧中的PLA-PEG@100mg/mL。 [0373] Solution 4: the burning of dichloromethane PLA-PEG @ 100mg / mL. 通过将化A-PEG溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by of A-PEG was dissolved in dichloromethane pure calcining.

[0374] 溶液5:在lOOmM的pH 8的憐酸盐缓冲液中的聚乙締醇@50mg/mL。 [0374] Solution 5: pity in acetate buffer pH lOOmM 8 in association polyvinyl alcohol @ 50mg / mL.

[037引首先制备第一油包水乳液。 [037 primer is first prepared a first water in oil emulsions. 通过将溶液1 ( 0.2 m L)、溶液2 ( 0 . 1 5 m L)、溶液3 (0.75mL)、W及溶液4(0.25mL)合并在一个小的压力管中并且使用布兰森数字超声波仪250 在50%振幅下进行声处理40秒来制备W1/01。 By the solution 1 (0.2 m L), a solution of 2 (0. 1 5 m L), a solution of 3 (0.75mL), W and a solution of 4 (0.25mL) were combined in a small pressure tube and using a digital Branson Sonifier 250 for 40 seconds sonication prepared W1 / 01 at 50% amplitude. 然后通过将溶液5(3.0mL)与第一W1/01乳液合并、满旋10秒、并且使用布兰森数字超声波仪250在30%振幅下进行声处理60秒来制备第二乳液(W1/01/W2)。 Then the solution was 5 (3.0mL) and the first W1 / 01 emulsion were combined, spin full 10 seconds, and using the Branson Digital Sonifier 250 at 30% amplitude for 60 seconds sonication prepared second emulsion (W1 / 01 / W2).

[0376] 将该W1/01/W2乳液添加到含有70mM的pH如溝酸盐缓冲溶液(30mL)的烧杯中并且在室溫下揽拌2小时W允许二氯甲烧蒸发并且允许纳米载体形成。 [0376] The added W1 / 01 / W2 emulsion to the beaker containing the groove as pH 70mM acetate buffer solution (30mL) of W and embrace for 2 hours at room temperature, allowing to burn methylene chloride was evaporated and allowed to form the nanocarrier . 通过W下方式来洗涂一部分纳米载体:将纳米载体悬浮液转移至一个离屯、管中,并且W75,600 Xg和4°C离屯、35分钟、去除上清液、并且将沉淀再悬浮于憐酸盐缓冲盐水中。 By way of a portion of the W-coated to wash the nanocarrier: The nanocarrier suspension was transferred to a village from the tube, and W75,600 Xg and 4 ° C from the village, 35 minutes, the supernatant was removed, and the pellet was resuspended pity salt in buffered saline. 重复该洗涂程序,并且将沉淀再悬浮于憐酸盐缓冲盐水中W用于约lOmg/mL的最终的纳米载体分散液。 The wash-coating procedure was repeated, and the pellet was resuspended in acetate buffer saline W pity for final nanocarriers of about lOmg / mL dispersion.

[0377] 通过动态光散射来确定纳米载体大小。 [0377] to determine the size of the nanocarrier by dynamic light scattering. 通过HPLC分析来确定在纳米载体中的肤和辛伐他汀的量。 To determine the amount of skin and simvastatin in the nanocarrier by HPLC analysis. 通过重量法来确定每mL悬浮液的总的干燥纳米载体质量。 Determining a total dry mass per mL nanocarrier suspension by the gravimetric method.

[037引 [037 Cited

Figure CN105999295AD00641

[0379] 体内给予1 [0379] 1 in vivo administration

[0380] 收获来自B6. Cg-Tg (TcraTcrb) 425Cbn/J (0TII)和C57BL/6 (B6)小鼠的脾脏、机械解离并且单独过滤通过70μΜ筛W产生单细胞悬浮液。 [0380] harvested from B6. Mouse spleen Cg-Tg (TcraTcrb) 425Cbn / J (0TII) and C57BL / 6 (B6), mechanically dissociated and individually produce a single cell suspension was filtered through a sieve 70μΜ W. 然后W2步骤的方法提取纯化的CD4+ CD25-细胞。 The method then steps W2, purified CD4 + CD25- cells. 使用美天旋生物技术公司(Miltenyi Biotec)的AutoMACS磁性细胞分选仪将脾脏细胞首先用CD4+T-细胞分离试剂盒II进行标记,并且未标记的部分用CD25耗尽试剂盒耗尽CD25+细胞。 US-day using a spin Biotech (Miltenyi Biotec) in the AutoMACS magnetic cell sorter The spleen cells were first treated with CD4 + T- cell isolation kit II labeled and unlabeled CD25 depleted portion by depletion of CD25 + cells kit . 纯化的B6细胞在与纯化的0ΤΠ 细胞W等浓度混合之前用细胞内染料簇基巧光素班巧酷亚胺醋(CFSE)进行染色。 Clever cluster group voxel light dye classes Qiao cool imine vinegar (of CFSE) cells stained with purified B6 intracellular concentration prior to mixing with the purified 0ΤΠ cell W or the like. 然后将它们静脉(iv)注射到B6.S化-P化rc^BoyAi (CD45.1)受体小鼠中。 They were then intravenously (iv) injected to B6.S of -P of rc ^ BoyAi (CD45.1) in recipient mice.

[0381] 第二天,用祀向的致耐受性合成疫苗颗粒(t2SVP)处理受体CD45.1小鼠。 [0381] The next day, treatment of the recipient mice were treated with Si to CD45.1 tolerogenic vaccine synthetic particles (t2SVP). 它们负载有卵白蛋白肤(323-339) (OVA 323^339)、雷帕霉素(Rapa)和/或辛伐他汀(Simva)的组合并且皮下地(SC)给予。 They skin loaded with ovalbumin (323-339) (OVA 323 ^ 339), the combination of rapamycin (Rapa) and / or simvastatin (Simva) and subcutaneously (SC) administration.

[0382] 注射构成一个致耐受性处理,并且随后进行另外4次注射,每次注射之间间隔2周。 [0382] Injection constituting a tolerogenic treatment, and then for another 4 injections at 2 week intervals between each injection. 在治疗计划完成之后,将受体CD45.1动物杀死并且收获它们的脾脏和順淋己结、机械解离并且单独过滤通过70ιχΜ筛W产生单细胞悬浮液。 After the treatment plan is completed, the animals were sacrificed and the recipient CD45.1 their spleens harvested cis lymph node hexyl, mechanical dissociation and individually produce a single cell suspension was filtered through a sieve 70ιχΜ W. 通过用RBC裂解缓冲液解育来使脾脏细胞耗尽红血细胞(RBC)(干细胞技术(Stem Cell Technologies))并且对脾脏和淋己结二者都进行细胞计数。 By incubation with a lysis buffer solution used to make the RBC spleen cells depleted of red blood cells (RBC) (Stem Cell Technology (Stem Cell Technologies)) and to have both the spleen and lymph node cells were counted for.

[0383] 将脾脏或淋己结细胞在增补有lOU/ml IL-2的CM(完全培养基)中培养、在96孔圆底(RB)培养板中W0.3 X 106细胞/孔用0PII再刺激、并且在37°C、5%C02下解育。 [0383] The spleen or lymph node cells had supplemented with lOU / ml CM IL-2 (complete medium) culture, culture plates W0.3 X 106 cells / well in 96 well round bottom (RB) with 0PII restimulation, and at 37 ° C, incubated under 5% C02 solution. 细胞在第2 天分裂并且在第5天被收获。 Cell division on day 2 and day 5 was harvested. 收集上清液并且冷冻,同时对细胞进行染色W便通过流式细胞术进行表型分析。 The supernatant was collected and frozen, while the cells were stained W will perform phenotypic analysis by flow cytometry. 运些细胞在碧迪公司的化CS化nto流式细胞仪上进行分析。 These cells were shipped in Becton Dickinson's based on the analysis of CS nto flow cytometry.

[0384] 体内给予2 [0384] 2 administered in vivo

[03 化]收获来自B6. Cg-Tg (TcraTcrb) 425Cbn/J (0TII)和C57BL/6 (B6)小鼠的脾脏、机械解离并且单独过滤通过70μΜ筛W产生单细胞悬浮液。 [Of 03] were harvested from B6. Cg-Tg (TcraTcrb) 425Cbn / J (0TII) and C57BL / 6 (B6) mouse spleen, mechanical dissociation and individually produce a single cell suspension was filtered through a sieve 70μΜ W. 然后使用美天旋生物技术公司AutoMACS磁性细胞分选仪W2步骤的方法提取纯化的CD4+CD25-细胞。 US days then spin Biotech AutoMACS magnetic cell sorter method step W2, purified CD4 + CD25- cells. 脾脏细胞使用美天旋的CD4+T-细胞分离试剂盒II来标记。 US-day spleen cells using the spin CD4 + T- cell isolation kit II marked. 未标记的CD4+T-细胞部分然后用CD25耗尽试剂盒耗尽CD25+细胞。 Unlabeled CD4 + T- cells were then depleted portion kit depleted CD25 + cells CD25. 然后,来自B6小鼠的纯化的CD4细胞在与纯化的0ΤΠ 细胞W等浓度混合之前用细胞内染料簇基巧光素班巧酷亚胺醋(CFSE)进行染色。 Then, purified CD4 cells from B6 mice were stained dye classes cluster group voxel Qiao Qiao light cool imine vinegar (of CFSE) with the intracellular concentration prior to mixing with the purified 0ΤΠ cell W or the like. 然后将它们静脉(iv)注射到B6. S化-P 化rc^/BoyAi (CD45.1)受体小鼠中。 They were then intravenously (iv) injected into B6. S -P technology of rc ^ / BoyAi (CD45.1) in recipient mice.

[0386] 第二天,用祀向的致耐受性合成疫苗颗粒处理受体CD45.1小鼠。 [0386] The next day, treatment of the recipient mice with induced CD45.1 Si particles to the synthetic vaccine resistance. 它们包含卵白蛋白肤(323-339) (OVA323-339 )、雷帕霉素(Rapa似及辛伐他汀(Simva)的组合并且皮下地(SC)或静脉内地(iv)给予。 They contain skin ovalbumin (323-339) (OVA323-339), the combination of rapamycin (Rapa like simvastatin (Simva) and subcutaneously (SC) or intravenously (iv) administration.

[0387] 在治疗计划完成之后,将受体CD45.1动物杀死并且收获它们的脾脏和順淋己结、 机械解离并且单独过滤通过7〇ΐ^Μ筛W产生单细胞悬浮液。 [0387] After the treatment plan is completed, the animals were sacrificed and the recipient CD45.1 their spleens harvested cis lymph node hexyl, mechanical dissociation and individually produce a single cell suspension was filtered through a sieve 7〇ΐ ^ [mu] W. 通过与RBC裂解缓冲液结合来使脾脏细胞耗尽红血细胞(RBC)(干细胞技术)并且对脾脏和淋己结二者都进行细胞计数。 Depleted spleen cells to make red blood cells (RBC) (Stem Cell Technologies) and has on both the spleen and lymph node cells were counted by both binding and RBC lysis buffer.

[0388]将脾脏或淋己结细胞在增补有lOU/ml化-2的CM中培养、在96孔圆底(RB)培养板中W0.3X106细胞/孔用ΙμΜ 0PII再刺激、并且在37°C、5%C02下解育。 [0388] The spleen or lymph node cells had supplemented with lOU / ml of culture in CM -2, culture plates W0.3X106 cells / well with ΙμΜ 0PII restimulated in 96 well round bottom (the RB), and 37 ° C, under 5% C02 sterile solutions. 细胞在第2天分裂并且在第5天被收获。 Cell division on day 2 and day 5 was harvested. 收集上清液并且冷冻,同时对细胞进行染色W便通过流式细胞术进行表型分析。 The supernatant was collected and frozen, while the cells were stained W will perform phenotypic analysis by flow cytometry. 运些细胞在碧迪公司的化CS化nto流式细胞仪上进行分析。 These cells were shipped in Becton Dickinson's based on the analysis of CS nto flow cytometry.

[0;3例结果 [0; 3 cases Results

[0390]运些结果示于图2和3(免疫抑制剂1:雷帕霉素;免疫抑制剂2:辛伐他汀)。 [0390] These operation results are shown in Figures 2 and 3 (1 immunosuppressants: rapamycin; immunosuppressant 2: simvastatin). 运些图显示了体内效应并且证明,与单独使用抗原或包含抗原而有和没有免疫刺激分子的合成纳米载体相比,使用包含抗原和免疫抑制剂的合成纳米载体使效应免疫细胞的数目降低。 These figures show a transport and demonstrated in vivo effects, alone or with the use of an antigen with and without antigen comprising an immunostimulatory synthetic nanocarriers molecules compared to synthetic nanocarriers containing antigen and immunosuppressant effect that the number of immune cells is reduced. 图3 还证明与仅包含抗原的合成纳米载体相比,使用包含抗原和免疫抑制剂的合成纳米载体使表达化xP3的百分数增加。 3 also demonstrated as compared with synthetic nanocarriers comprise only an antigen, comprising an antigen using synthetic nanocarriers immunosuppressants and expressing the percentage increase of xP3.

[0巧1] 实例17:评定具有抗原和免疫抑制剂的纳米载体的作用 [0 Qiao 1] Example 17: Evaluation of the role of the nanocarrier with antigens and immunogens inhibitor

[0392] 纳米载体 [0392] nanocarrier

[0393] 从己亨美洲公司(柏市街3132号,托伦斯,加利福尼亚州90505,区号#4065609)购买卵白蛋白肤323-339,即一种已知为卵白蛋白的T和B细胞表位的17氨基酸肤。 [0393] purchase ovalbumin own skin 323-339 from Hang Americas (No. 3132 Kashiwa Street, Torrance, California 90505, area code # 4065609), that is known as the ovalbumin T and B cell epitopes 17 amino acids skin. 从SurModics制药公司(汤姆马下路756号,伯明翰,亚拉己马州35211;产品代码7525化G 7A) 购买丙交醋:乙交醋比为3:1并且特性粘度为0.75化/g的化GA。 Pharmaceuticals from SurModics (Tang Muma drop 756, Birmingham, hexyl Maryland 35211; product code 7525 of G 7A) for later vinegar lactide: glycolide ratio of acetate 3: 1 and an intrinsic viscosity of 0.75 / g, of GA. 合成了PEG嵌段大约为5, OOODa并且化A嵌段大约为20,000Da的化A-PEG嵌段共聚物。 Synthesis of the PEG blocks is about 5, OOODa and A blocks of A-PEG of about 20,000Da of the block copolymer. 从EMD化学品公司(产品编号1.41350.1001)购买聚乙締醇(85%-89%水解的)。 Polyvinyl alcohol for later association from the EMD Chemicals (Catalog No .: 1.41350.1001) (85% -89% hydrolyzed).

[0394] 如下制备溶液:溶液1:在稀释的盐酸水溶液中的卵白蛋白肤323-339@20mg/mL。 [0394] A solution was prepared as follows: Solution 1: the diluted aqueous hydrochloric acid skin ovalbumin 323-339 @ 20mg / mL. 通过在室溫下将卵白蛋白肤溶解在0.13M盐酸溶液中而制备该溶液。 The solution prepared in 0.13M hydrochloric acid solution by the skin of ovalbumin dissolved at room temperature. 溶液2:在二氯甲烧中的化GA@1 OOmg/mL。 Solution 2: In the burning of dichloromethane GA @ 1 OOmg / mL. 通过将化GA溶解在纯二氯甲烧中而制备该溶液。 The solution of GA is prepared by dissolving the pure dichloromethane calcining. 溶液3 :在二氯甲烧中的PLA-PEG@ 1 OOmg/mL。 Solution 3: In the burning of dichloromethane PLA-PEG @ 1 OOmg / mL. 通过将PLA-PEG溶解在纯二氯甲烧中而制备该溶液。 The solution is prepared by dissolving PLA-PEG pure dichloromethane calcining. 溶液4:在1 OOmM的pH 8的憐酸盐缓冲液中的聚乙締醇@50mg/mL。 Solution 4: 1 OOmM pity salt buffer of pH 8 in association polyvinyl alcohol @ 50mg / mL. 首先制备第一油包水乳液。 First preparing a first water in oil emulsions. 通过将溶液1 (0.2mL)、溶液2(0.75mL)、W及溶液3(0.25mL)合并在一个小的压力管中并且使用布兰森数字超声波仪250在50%振幅下进行声处理40秒来制备W1/01。 By the solution 1 (0.2mL), a solution of 2 (0.75mL), W and a solution (0.25 mL of) 3 were combined in a small pressure tube and using Branson Digital Sonifier 250 at 50% amplitude sonication 40 seconds to prepare W1 / 01. 然后通过将溶液4(3.OmL)与第一W1/01乳液合并、满旋10秒、并且使用布兰森数字超声波仪250在30%振幅下进行声处理60秒来制备第二乳液(W1/01/W2)。 The solution was then passed through 4 (3.OmL) and the first W1 / 01 emulsion were combined, spin full 10 seconds, and using the Branson Digital Sonifier 250 acoustically at 30% amplitude for 60 seconds to prepare a second emulsion (W1 / 01 / W2).

[0395] 将该W1/01/W2乳液添加到含有70mM的pH如溝酸盐缓冲溶液(30mL)的烧杯中并且在室溫下揽拌2小时W允许二氯甲烧蒸发并且允许纳米载体形成。 [0395] The added W1 / 01 / W2 emulsion to the beaker containing the groove as pH 70mM acetate buffer solution (30mL) of W and embrace for 2 hours at room temperature, allowing to burn methylene chloride was evaporated and allowed to form the nanocarrier . 通过W下方式来洗涂一部分纳米载体:将纳米载体悬浮液转移至一个离屯、管中,并且W75,600Xg和4°C离屯、35分钟、去除上清液、并且将沉淀再悬浮于憐酸盐缓冲盐水中。 By way of a portion of the W-coated to wash the nanocarrier: The nanocarrier suspension was transferred to a village from the tube, and W75,600Xg and 4 ° C from the village, 35 minutes, the supernatant was removed, and the pellet was resuspended in Rei-buffered saline. 重复该洗涂程序,并且将沉淀再悬浮于憐酸盐缓冲盐水中W用于约lOmg/mL的最终的纳米载体分散液。 The wash-coating procedure was repeated, and the pellet was resuspended in acetate buffer saline W pity for final nanocarriers of about lOmg / mL dispersion.

[0396] 通过动态光散射来确定纳米载体大小。 [0396] to determine the size of the nanocarrier by dynamic light scattering. 通过HPLC分析来确定在纳米载体中的肤的量。 To determine the amount of skin in the nanocarrier by HPLC analysis. 通过重量法来确定每mL悬浮液的总的干燥纳米载体质量。 Determining a total dry mass per mL nanocarrier suspension by the gravimetric method.

[0397] [0397]

Figure CN105999295AD00661

[0398] 从己亨美洲公司(柏市街3132号,托伦斯,加利福尼亚州90505,区号#4065609)购买卵白蛋白肤323-339,即一种已知为卵白蛋白的T和B细胞表位的17氨基酸肤。 [0398] purchase ovalbumin own skin 323-339 from Hang Americas (No. 3132 Kashiwa Street, Torrance, California 90505, area code # 4065609), that is known as the ovalbumin T and B cell epitopes 17 amino acids skin. 从化学品公司(威尔逊街185号,弗雷明汉,马萨诸塞州01702;产品目录#31017)购买雷帕霉素。 Buy rapamycin; from the chemical company (Catalog # 31017 185 Wilson Street, Framingham, MA 01702). 从SurModics制药公司(汤姆马下路756号,伯明翰,亚拉己马州35211;产品代码7525化G 7Α) 购买丙交醋:乙交醋比为3:1并且特性粘度为0.75化/g的化GA。 Pharmaceuticals from SurModics (Tang Muma drop 756, Birmingham, hexyl Maryland 35211; product code 7525 of G 7Α) vinegar later lactide: glycolide ratio of acetate 3: 1 and an intrinsic viscosity of 0.75 / g, of GA. 合成了PEG嵌段大约为5, OOODa并且化A嵌段大约为20,000Da的化A-PEG嵌段共聚物。 Synthesis of the PEG blocks is about 5, OOODa and A blocks of A-PEG of about 20,000Da of the block copolymer. 从EMD化学品公司(产品编号1.41350.1001)购买聚乙締醇(85%-89%水解的)。 Polyvinyl alcohol for later association from the EMD Chemicals (Catalog No .: 1.41350.1001) (85% -89% hydrolyzed).

[0399] 如下制备溶液:溶液1:在稀释的盐酸水溶液中的卵白蛋白肤323-339@20mg/mL。 [0399] A solution was prepared as follows: Solution 1: the diluted aqueous hydrochloric acid skin ovalbumin 323-339 @ 20mg / mL. 通过在室溫下将卵白蛋白肤溶解在0.13M盐酸溶液中而制备该溶液。 The solution prepared in 0.13M hydrochloric acid solution by the skin of ovalbumin dissolved at room temperature. 溶液2:在二氯甲烧中的雷帕霉素@50mg/mL。 Solution 2: Burning in dichloromethane rapamycin @ 50mg / mL. 通过将雷帕霉素溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by dissolving the pure rapamycin dichloromethane calcining. 溶液3:在二氯甲烧中的PLGA@1 OOmg/mL。 Solution 3: In the burning of dichloromethane PLGA @ 1 OOmg / mL. 通过将PLGA溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by dissolving the PLGA in neat dichloromethane calcining. 溶液4:在二氯甲烧中的化A-PEG@1 OOmg/mL。 Solution 4: the burning of dichloromethane of A-PEG @ 1 OOmg / mL. 通过将化A-PEG溶解在纯二氯甲烧中而制备该溶液。 The solution was prepared by of A-PEG was dissolved in dichloromethane pure calcining. 溶液5 :在lOOmM的pH 8的憐酸盐缓冲液中的聚乙締醇@50mg/mL。 Solution 5: pity in acetate buffer pH lOOmM 8 in association polyvinyl alcohol @ 50mg / mL.

[0400] 首先制备第一油包水乳液。 [0400] First preparing a first water in oil emulsions. 通过将溶液l(〇.2mL)、溶液2(0.2mL)、溶液3(0.75mL)、 W及溶液4(0.25mL)合并在一个小的压力管中并且使用布兰森数字超声波仪250在50%振幅下进行声处理40秒来制备W1/01。 By the solution L (〇.2mL), a solution of 2 (0.2mL), a solution of 3 (0.75mL), W and a solution of 4 (0.25mL) were combined in a small pressure tube and using Branson Digital Sonifier 250 sonicated at 50% amplitude for 40 seconds to prepare W1 / 01. 然后通过将溶液5(3.0mL)与第一W1/01乳液合并、满旋10秒、并且使用布兰森数字超声波仪250在30%振幅下进行声处理60秒来制备第二乳液(W1/01/W2)。 Then the solution was 5 (3.0mL) and the first W1 / 01 emulsion were combined, spin full 10 seconds, and using the Branson Digital Sonifier 250 at 30% amplitude for 60 seconds sonication prepared second emulsion (W1 / 01 / W2).

[0401 ]将该W1/01/W2乳液添加到含有70mM的pH如溝酸盐缓冲溶液(30mL)的烧杯中并且在室溫下揽拌2小时W允许二氯甲烧蒸发并且允许纳米载体形成。 [0401] The added W1 / 01 / W2 emulsion to the beaker containing the groove as pH 70mM acetate buffer solution (30mL) of W and embrace for 2 hours at room temperature, allowing to burn methylene chloride was evaporated and allowed to form the nanocarrier . 通过W下方式来洗涂一部分纳米载体:将纳米载体悬浮液转移至一个离屯、管中,并且W21,000Xg和4°C离屯、45分钟、去除上清液、并且将沉淀再悬浮于憐酸盐缓冲盐水中。 By way of a portion of the W-coated to wash the nanocarrier: The nanocarrier suspension was transferred to a village from the tube, and W21,000Xg and 4 ° C from the village, 45 minutes, the supernatant was removed, and the pellet was resuspended in Rei-buffered saline. 重复该洗涂程序,并且将沉淀再悬浮于憐酸盐缓冲盐水中W用于约lOmg/mL的最终的纳米载体分散液。 The wash-coating procedure was repeated, and the pellet was resuspended in acetate buffer saline W pity for final nanocarriers of about lOmg / mL dispersion.

[0402] 通过动态光散射来确定纳米载体大小。 [0402] to determine the size of the nanocarrier by dynamic light scattering. 通过HPLC分析来确定在纳米载体中的肤和雷帕霉素的量。 Skin and to determine the amount of rapamycin in the nanocarrier by HPLC analysis. 通过重量法来确定每mL悬浮液的总的干燥纳米载体质量。 Determining a total dry mass per mL nanocarrier suspension by the gravimetric method.

[0403] [0403]

Figure CN105999295AD00662

[0404] 通过动态光散射(DLS)获得合成纳米载体尺寸的测量值。 [0404] obtaining measurements synthetic nanocarriers sized by dynamic light scattering (DLS). 将合成纳米载体的悬浮液用纯化水稀释到W达到近似0.01至O.lmg/mL的最终的合成纳米载体悬浮液浓度。 The aqueous suspension was purified synthetic nanocarriers W to reach a final dilution of synthetic nanocarriers suspension concentration approximately 0.01 to O.lmg / mL of. 在一个适合的比色皿中直接制备稀释的悬浮液W用于化S分析。 W suspensions prepared directly diluted in a suitable cuvette for analysis of S. 然后将该比色皿置于布鲁克海文仪器公司(Brookhaven Instruments Co;rp. )ZetaPALS中,允许其平衡至25°C,并且然后扫描充分的时间W在介质的粘度和样品的折射率的适当输入的基础上获得稳定且可再现的分布。 The cuvette is then placed in Brookhaven Instruments Corporation (Brookhaven Instruments Co;. Rp) ZetaPALS, allowing it to equilibrate to 25 ° C, and then the scan time W sufficient viscosity and suitable refractive index of the sample medium stable distribution on the basis of input and reproducible. 然后报告有效直径,或该分布的平均值。 Then reports the effective diameter, or the average of the distribution.

[0405] 免疫 [0405] immunity

[0406] 将纳米载体融化并平衡。 [0406] The nanocarriers melted and equilibrated. 初始稀释液构成一个lOx储备溶液,并且进一步在OVA323-339中稀释到l(K)μg/ml的浓度或lx溶液。 The initial dilution constitute a lOx stock solution and further diluted to a concentration of solution or lx l (K) μg / ml in OVA323-339. 将运个lx溶液用于W每静脉内注射按20化1 进行注射。 The solution was used to transport a lx every intravenous injection of 1 W at 20 for injection. 将动物用OVA蛋白(OVA)免疫并且用OVA323-339肤处理。 Animals treated with OVA323-339 skin with OVA protein (OVA) and immunized. 免疫途径如下:1化g的0VA+ 4mg明抓(Alum),腹膜内,每只Ba化/C初次接受免疫的雌性小鼠40化1。 Immunization route is as follows: 1 g of a clear grasp 0VA + 4mg (Alum were), intraperitoneal, each of the Ba / C female mice were immunized naïve of 40 1. 实验组各自由5只动物组成。 Experimental groups will consist of five animals. 脾细胞用抗原使用CFSE或CT0重新刺激,W便测定抗原特异性增殖的量。 CFSE or using spleen cells re-stimulated with antigen CT0, the amount of antigen-specific proliferation assay W will.

[0407] 具体免疫细胞类型的水平 [0407] Specific types of immune cell levels

[040引使用Flow化软件分析FCS文件。 [040 citation analysis FCS file using Flow software. 排除7AAD阳性细胞(一种标记死细胞的核染色)并且将依赖于CD4、CD8、Gr-l、F4/80、B220、TC肺W及CDnb表达的细胞形态定量。 7AAD negative-positive cells (one labeled nuclear staining dead cells) and dependent on CD4, CD8, Gr-l, F4 / 80, Morphometric cells expressing B220, TC and lung W CDnb.

[0409] 用于T细胞子集的口控策略一7AAD-F4/80-GR-1-TC 肺+CD4+/-CD8+/- [0409] Policy control port for a T cell subset 7AAD-F4 / 80-GR-1-TC + CD4 +/- CD8 +/- lung

[0410] 分裂CD4巧细胞和CD8巧细胞%的测定 [0410] Qiao division CD4 cells and CD8 cells Qiao% assay

[0411] 借助流式细胞术计算卵白蛋白反应性CD4+T细胞和CD8巧细胞的频率。 [0411] Flow cytometry is calculated by means of ovalbumin frequency of CD8 + T cells and CD4 cell reactivity clever. 将来自实验动物的脾细胞用一种适用于长期细胞标记的硫醇反应性巧光探针CFSE染色,并且在完全培养基中在37 °C、5 % C〇2下用卵白蛋白培养3天。 Spleen cells from the experimental animal in a suitable thiol-reactive long cell marker clever optical probe CFSE staining, and in complete medium at 37 ° C, 5% under C〇2 cultured for 3 days with ovalbumin . 在第3天,将细胞洗涂、用抗CD16/32抗体阻断并接着用对TCR CD4和CD8a特异的共辆抗体染色。 On day 3, cells were washed coated, blocked with CD16 / 32 antibody and vehicle were then stained with antibodies specific for the TCR CD4 and CD8a. 通过比较差异性CFSE染色评定TCR+CD4或TCR+CD8a+脾细胞的增殖。 TCR CD4 or TCR CD8a + + + splenocytes proliferation assessed by CFSE staining comparing differences.

[0412] 结果 [0412] results

[0413]图4和图5证明了运些纳米载体在动物模型中的有效性。 [0413] Figures 4 and 5 demonstrate the effectiveness of the nanocarrier in operation some animal models. 具体来说,图4证明了来自用包含OVA323-339和免疫抑制剂的合成纳米载体治疗的动物受试者的灌洗样品中CD4巧细胞和CD8巧细胞的数目减少。 Specifically, Figure 4 demonstrates a reduction in the number of animals lavage sample from the subject treated with synthetic nanocarrier comprising OVA323-339 and immunosuppressants clever in CD4 cells and CD8 cells clever. 图5显示了使用包含免疫抑制剂和OVA肤的合成纳米载体的给予使分裂CD4巧细胞和CD8巧细胞的百分数降低。 Figure 5 shows a synthetic nanocarrier comprising an immunosuppressant and a skin of OVA administered so clever division CD4 and CD8 cell percentage decreased clever.

[0414] 实例18:评定具有抗原和免疫抑制剂的纳米载体的作用(预示的) [0414] Example 18: Evaluation of the role of antigenic and immunosuppressive agents nanocarriers (Prophetic)

[041引纳米载体 [041 cited nanocarrier

[0416] 类似于W上实例(实例17)使用Μ肥I类限制性肤SINF邸L(沈Q ID N0:944)代替卵白蛋白肤来制备纳米载体。 [0416] W similar to example (example 17) using the fertilizer class I restricted Μ skin SINF Di L (Shen Q ID N0: 944) was prepared in place of ovalbumin nanocarriers skin.

[0417] 免疫 [0417] immunity

[0418] 将纳米载体融化并平衡。 [0418] The nanocarriers melted and equilibrated. 初始稀释液构成一个lOx储备溶液,并且进一步在肤中稀释到l(K)μg/ml的浓度或lx溶液。 The initial dilution constitute a lOx stock solution and further diluted to a concentration of solution or lx l (K) μg / ml in the skin. 将运个lx溶液用于W每静脉内注射按20化1进行注射。 The solution was used to transport a lx every intravenous injection of 1 W at 20 for injection. 将动物用包含该肤的蛋白质免疫并且用肤处理。 The animals immunized with protein containing the skin and skin treatment. 免疫途径如下:l〇yg的肤+4mg明抓(A1皿),腹膜内,每只Ba化/C初次接受免疫的(immunologically.n幻机6)雌性小鼠40化1。 Immunization route is as follows: The skin l〇yg + 4mg next catch (A1 dish), intraperitoneal, each of the Ba / C immunized naïve (immunologically.n magic machine 6) female mice of 40 1. 实验组各自由5 只动物组成。 Experimental groups will consist of five animals. 脾细胞用抗原使用CFSE或CT0重新刺激,W便测定抗原特异性增殖的量。 CFSE or using spleen cells re-stimulated with antigen CT0, the amount of antigen-specific proliferation assay W will.

[0419] 具体免疫细胞类型的水平 [0419] Specific types of immune cell levels

[0420] 使用Flow化软件分析FCS文件。 [0420] FCS file using Flow analysis software. 排除7AAD阳性细胞(一种标记死细胞的核染料)并且将依赖于CD4、CD8、Gr-l、F4/80、B220、TC肺W及CDnb表达的细胞形态定量。 7AAD negative-positive cells (one labeled nuclear stain dead cells) and dependent on CD4, CD8, Gr-l, F4 / 80, Morphometric cells expressing B220, TC and lung W CDnb.

[0421] 用于T细胞子集的口控策略一7AAD-F4/80-GR-1-TC 肺+CD4+/-CD8+/- [0421] Policy control port for a T cell subset 7AAD-F4 / 80-GR-1-TC + CD4 +/- CD8 +/- lung

[0422] 分裂CD4巧细胞和CD8巧细胞%的测定 [0422] Qiao division CD4 cells and CD8 cells Qiao% assay

[0423]借助流式细胞术计算卵白蛋白反应性CD4+T细胞和CD8+T细胞的频率。 [0423] Flow cytometry is calculated by means of ovalbumin frequency of CD8 + T cells and CD4 + T cell reactivity. 将来自实验动物的脾细胞用一种适用于长期细胞标记的硫醇反应性巧光探针CFSE染色,并且在完全培养基中在37 °C、5 % C〇2下用卵白蛋白培养3天。 Spleen cells from the experimental animal in a suitable thiol-reactive long cell marker clever optical probe CFSE staining, and in complete medium at 37 ° C, 5% under C〇2 cultured for 3 days with ovalbumin . 在第3天,将细胞洗涂、用抗CD16/32抗体阻断并接着用对TCR CD4和CD8a特异的共辆抗体染色。 On day 3, cells were washed coated, blocked with CD16 / 32 antibody and vehicle were then stained with antibodies specific for the TCR CD4 and CD8a. 通过比较差异性CFSE染色评定TCR+CD4或TCR+CD8a+脾细胞的增殖。 TCR CD4 or TCR CD8a + + + splenocytes proliferation assessed by CFSE staining comparing differences.

Claims (51)

1. 一种方法,包括: 根据一种治疗方案向受试者给予一种组合物,该治疗方案先前已经显示降低一个或多个测试受试者中的抗原特异性效应性T细胞的数目或活性; 其中,该组合物包含: (i) 第一群体的合成纳米载体,这些第一群体的合成纳米载体与免疫抑制剂偶联,和(ii) 第二群体的合成纳米载体,这些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联。 1. A method, comprising: administering to the subject a composition according to a therapeutic regimen, the treatment regimen to reduce the number of the one or more previous test subject antigen-specific effector T cells have been shown or activity; wherein the composition comprises: (i) a first population of synthetic nanocarriers, synthetic nanocarriers immunosuppressant first coupling these groups, and synthetic nanocarriers (ii) a second population, the second synthetic nanocarriers groups with MHC class I-restricted epitope of an antigen and / or MHC class II restricted epitopes conjugated.
2. -种方法,包括: 通过向一个或多个测试受试者给予一种组合物来降低该一个或多个测试受试者中的抗原特异性效应性T细胞的数目或活性,该组合物包含: (i) 第一群体的合成纳米载体,这些第一群体的合成纳米载体与免疫抑制剂偶联,和(ii) 第二群体的合成纳米载体,这些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联。 2. - method, comprising: reducing the number or activity of one or more of the test subject antigen-specific effector T cells by administering a composition to one or more of the test subject, the composition comprising: (i) a first population of synthetic nanocarriers, synthetic nanocarriers immunosuppressant first coupling these groups, and synthetic nanocarriers (ii) a second population, second population of these synthetic nanocarriers and an antigen MHC class I restricted epitope and / or MHC class II restricted epitopes conjugated.
3. -种方法,包括: 向受试者给予一种组合物,该组合物包含: (i) 第一群体的合成纳米载体,这些第一群体的合成纳米载体与免疫抑制剂偶联,和(ii) 第二群体的合成纳米载体,这些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联; 其中,该组合物处于有效降低抗原特异性效应性T细胞的数目或活性的量。 3. The - method, comprising: administering a composition to a subject, the composition comprising: (i) a first population of synthetic nanocarriers, synthetic nanocarriers immunosuppressant coupling these first groups, and synthetic nanocarriers (ii) a second population, a second population of synthetic nanocarriers of these MHC class I-restricted epitopes of one antigen-restricted epitopes coupling and / or MHC II; wherein the composition is in amount or activity of the number of antigen-specific effector T cells effectively reduced.
4. 如权利要求1-3中任一项所述的方法,其中该第一群体和该第二群体是相同的群体。 4. A method according to any one of claims, wherein the first population and the second population are identical groups.
5. 如权利要求1-4中任一项所述的方法,其中该方法进一步包括提供或鉴定该受试者。 5. The method according to any one of claims 1-4, wherein the method further comprises providing or identifying the subject.
6. 如权利要求1-5中任一项所述的方法,其中该抗原是一种治疗性蛋白、一种自身抗原或一种过敏原,或与一种炎性疾病、一种自身免疫性疾病、器官或组织排斥或移植物抗宿主病相关。 6. A method as claimed in any one of claims, wherein the therapeutic protein is an antigen, a self antigen or an allergen or with an inflammatory disease, an autoimmune disease, organ or tissue rejection or graft-versus-host disease related.
7. 如权利要求1-6中任一项所述的方法,其中该方法进一步包括在给予该组合物之前和/或之后评定该受试者中的抗原特异性效应性T细胞的数目或活性。 7. A method as claimed in any one of claims 1-6, wherein the method further includes assessing the number or activity of the subject antigen-specific effector T cells prior to administration of the composition and / or after .
8. 如权利要求1-7中任一项所述的方法,其中该受试者具有或正处于具有一种炎性疾病、一种自身免疫性疾病、一种过敏症、器官或组织排斥或移植物抗宿主病的风险。 8. A method according to any one of claims 1-7, wherein the subject has or is in having an inflammatory disease, an autoimmune disease, an allergic disease, organ or tissue rejection, or the risk of graft-versus-host disease.
9. 如权利要求1-7中任一项所述的方法,其中该受试者已经经历或将要经历移植。 9. The method according to any one of the preceding claims, wherein the subject has undergone or will undergo transplantation.
10. 如权利要求1-7中任一项所述的方法,其中该受试者具有或正处于具有针对一种正被给予或将要被给予该受试者的治疗性蛋白的一种所不希望的免疫应答的风险。 10. The method according to any one of claims 1-7, wherein the subject has or is having a positive A positive against one is administered or is to be administered to the subject therapeutic protein are not We want to risk an immune response.
11. 如权利要求1-10中任一项所述的方法,其中向该受试者给予一个或多个维持剂量的包含这些第一群体和第二群体的合成纳米载体的组合物。 11. The method according to any one of claims 1-10 wherein the administering to the subject one or more maintenance dose comprises a first population and second population of these compositions synthetic nanocarriers.
12. 如权利要求1-11中任一项所述的方法,其中该给予是通过静脉内、腹膜内、透粘膜、 经口、皮下、肺部、鼻内、皮内或肌肉内给予进行的。 12. A method as claimed in any one of claims 1 to 11, wherein the administration is by intravenous, intraperitoneal, transmucosal, oral, subcutaneous, pulmonary, intranasal, transdermal or intramuscular administration for the .
13. 如权利要求1-11中任一项所述的方法,其中该给予是通过吸入或静脉内、皮下或透粘膜给予进行的。 13. The method according to any one of claims 1-11 wherein the administration is by inhalation or intravenous, subcutaneous or transmucosal administration performed.
14. 如权利要求1-13中任一项所述的方法,其中这些免疫抑制剂包含一种抑制素、一种mTOR抑制剂、一种TGF-β信号剂、一种皮质类固醇、一种线粒体功能抑制剂、一种P38抑制剂、 一种NF-κβ抑制剂、一种腺苷受体激动剂、一种前列腺素E2激动剂、一种磷酸二酯酶4抑制剂、一种HDAC抑制剂、或一种蛋白酶体抑制剂。 14. The method according to any one of claims 1 to 13, wherein the immunosuppressive agent comprises one statin, one mTOR inhibitor, a TGF-β signal reagent A corticosteroid, a mitochondrial function inhibitor, a P38 inhibitor An inhibitor of NF-κβ A adenosine receptor agonist, a prostaglandin E2 agonist A phosphodiesterase 4 inhibitor, a HDAC inhibitor or a protease inhibitor.
15. 如权利要求14所述的方法,其中该mTOR抑制剂是雷帕霉素。 15. The method according to claim 14, wherein the mTOR inhibitor is rapamycin.
16. 如权利要求1-15中任一项所述的方法,其中平均在这些第一和/或第二群体的合成纳米载体上的这些免疫抑制剂和/或表位的负载是在0.0001 %与50%之间(重量/重量)。 16. The method according to any one of 1-15 claims, wherein the immunosuppressive and / or average loads epitopes on these synthetic nanocarriers first and / or second population is at 0.0001% between 50% (wt / wt).
17. 如权利要求16所述的方法,其中平均在这些第一和/或第二群体的合成纳米载体上的这些免疫抑制剂和/或表位的负载是在0.1 %与10%之间(重量/重量)。 17. The method of claim 16, wherein the average of these immunosuppressants on synthetic nanocarriers These first and / or second groups and / or load epitope is between 0.1% and 10% ( weight / weight).
18. 如权利要求1-17中任一项所述的方法,其中该第一群体和/或第二群体的这些合成纳米载体包括脂质纳米颗粒、聚合物纳米颗粒、金属纳米颗粒、基于表面活性剂的乳液、树枝状化合物、巴基球、纳米线、病毒样颗粒、或肽或蛋白质颗粒。 18. The method as claimed in any one of claims 1-17, wherein the first population and / or the synthetic nanocarriers second population comprises lipid nanoparticles, polymeric nanoparticles, metal nanoparticles, based on surface the emulsion of the active agent, dendrimers, buckyballs, nanowires, virus-like particle, or a peptide or protein particles.
19. 如权利要求18所述的方法,其中该第一群体和/或第二群体的这些合成纳米载体包括脂质纳米颗粒。 19. The method as claimed in claim 18, wherein the first population and / or the synthetic nanocarriers second population comprises lipid nanoparticles.
20. 如权利要求19所述的方法,其中该第一群体和/或第二群体的这些合成纳米载体包括脂质体。 20. The method according to claim 19, wherein the first population and / or the synthetic nanocarriers comprises a second population of liposomes.
21. 如权利要求18所述的方法,其中该第一群体和/或第二群体的这些合成纳米载体包括金属纳米颗粒。 21. The method according to claim 18, wherein the first population and / or the synthetic nanocarriers second population comprises metal nanoparticles.
22. 如权利要求21所述的方法,其中这些金属纳米颗粒包括金纳米颗粒。 22. The method according to claim 21, wherein the metal nanoparticles comprise gold nanoparticles.
23. 如权利要求18所述的方法,其中该第一群体和/或第二群体的这些合成纳米载体包括聚合物纳米颗粒。 23. The method according to claim 18, wherein the first population and / or the synthetic nanocarriers comprises a second population of polymeric nanoparticles.
24. 如权利要求23所述的方法,其中该聚合物纳米颗粒包括聚合物,该聚合物是一种非甲氧基封端的普朗尼克聚合物。 24. The method according to claim 23, wherein the polymeric nanoparticles comprise a polymer, the polymer is a non-methoxy-terminated Pluronic polymers.
25. 如权利要求23或24所述的方法,其中这些聚合物纳米颗粒包括聚酯、与聚醚偶联的聚酯、聚氨基酸、聚碳酸酯、聚缩醛、聚缩酮、多糖、聚乙基噁唑啉或聚乙烯亚胺。 25. The method of claim 23 or claim 24, wherein the polymeric nanoparticles polyesters include polyesters, polyethers and coupled, polyamino acids, polycarbonates, polyacetals, ketals, polysaccharides, poly ethyl oxazoline or polyethyleneimine.
26. 如权利要求25所述的方法,其中该聚酯包括一种聚(乳酸)、聚(乙醇酸)、聚(乳酸-乙醇酸)共聚物、或聚己酸内酯。 26. The method according to claim 25, wherein the polyester comprises a poly (lactic acid), poly (glycolic acid), poly (lactic acid - glycolic acid) copolymer, or a polycaprolactone.
27. 如权利要求25或26所述的方法,其中这些聚合物纳米颗粒包括聚酯以及与聚醚偶联的聚酯。 27. The method of claim 25 or claim 26, wherein the polymeric nanoparticles comprise a polyester and a polyether polyester conjugated.
28. 如权利要求25-27中任一项所述的方法,其中该聚醚包括聚乙二醇或聚丙二醇。 28. The method according to any one of claims 25-27, wherein said polyether comprises polyethylene glycol or polypropylene glycol.
29. 如权利要求1-28中任一项所述的方法,其中使用对该第一和/或第二群体的这些合成纳米载体进行的动态光散射而获得的粒度分布的平均值是大于l〇〇nm的直径。 29. The method according to any one of claims 1 to 28, wherein the average particle size distribution using dynamic light scattering synthetic nanocarriers the first and / or second population will be obtained is greater than l 〇〇nm diameter.
30. 如权利要求29所述的方法,其中该直径大于150nm。 30. The method according to claim 29, wherein the diameter is greater than 150nm.
31. 如权利要求30所述的方法,其中该直径大于200nm。 31. The method according to claim 30, wherein the diameter is greater than 200nm.
32. 如权利要求31所述的方法,其中该直径大于250nm。 32. The method according to claim 31, wherein the diameter is greater than 250nm.
33. 如权利要求32所述的方法,其中该直径大于300nm。 33. The method according to claim 32, wherein the diameter is greater than 300nm.
34. 如权利要求1-33中任一项所述的方法,其中该第一群体和/或第二群体的这些合成纳米载体的长宽比大于1:1、1:1.2、1:1.5、1:2、1:3、1:5、1:7或1:10。 34. A method as claimed in any one of claims 1-33, wherein the aspect ratio of the first group and / or the synthetic nanocarriers second population is greater than 1: 1, 1: 1.2, 1: 1.5, 1: 2,1: 3,1: 5,1: 7 or 1:10.
35. 如权利要求1-34中任一项所述的方法,其中这些效应性T细胞是⑶4+和/或CD8+T细胞。 The method of any one of claims 1-34 as claimed in claim 35, wherein the effector T cells are ⑶4 + and / or CD8 + T cells.
36. 一种方法,包括: (i) 产生第一群体的合成纳米载体,这些第一群体的合成纳米载体与免疫抑制剂偶联, 并且(ii) 产生第二群体的合成纳米载体,这些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联;并且(iii) 评估这些第一和第二群体的合成纳米载体对抗原特异性效应性T细胞数目或活性的作用。 36. A method, comprising: (i) generating a first group of synthetic nanocarriers, synthetic nanocarriers immunosuppressant first coupling these groups, and (ii) to produce a second population of synthetic nanocarriers, which first synthetic nanocarriers two groups with MHC class I-restricted epitopes of an antigen and / or MHC class II restricted epitopes coupling; and (iii) evaluation of these synthetic nanocarriers first and second populations of antigen specific role of effector cell number or activity of T.
37. 如权利要求36所述的方法,其中该第一群体和第二群体是相同的群体。 37. The method according to claim 36, wherein the first population and the second population are identical groups.
38. 如权利要求36或37所述的方法,其中该方法进一步包括制造一种剂型,该剂型包含这些第一群体和第二群体的合成纳米载体。 The method of claim 36 or claim 37, wherein the method further comprises producing a dosage form comprising a first synthetic nanocarrier these groups and the second groups.
39. 如权利要求36-38中任一项所述的方法,其中该方法进一步包括制造一种包含这些第一群体和第二群体的合成纳米载体的组合物、或可供受试者给予的剂型。 39. A method as claimed in any one of claims 36-38, wherein the method further comprises producing a composition comprising a first population of these synthetic nanocarriers and second groups, or administered to a subject for formulations.
40. 如权利要求39所述的方法,其中该方法进一步包括在给予该组合物或剂型之前和/ 或之后评定该受试者中的抗原特异性效应性T细胞的数目或活性。 40. The method according to claim 39, wherein the method further includes assessing the number or activity of the subject antigen-specific effector T cells and / or before and after administration of the composition or dosage form.
41. 如权利要求36-40中任一项所述的方法,其中这些效应性T细胞是CD4+T细胞和/或⑶8+T细胞。 41. The method according to any one of claims 36-40, wherein the effector T cell is a CD4 + T cells and / or ⑶8 + T cells.
42. 如权利要求36-41中任一项所述的方法,其中产生的这些第一群体和第二群体的合成纳米载体是如在权利要求1-35中任一项所定义的。 42. The method according to any one of claims 36-41, these synthetic nanocarriers first population and second population is generated therein as defined in any one of 1-35 claims.
43. -种用于生产一种组合物或剂型的工艺,该工艺包括以下步骤: (i) 将第一群体的合成纳米载体与免疫抑制剂偶联; (ii) 将第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联,并且(iii) 评估这些第一和第二群体的合成纳米载体对抗原特异性效应性T细胞数目或活性的作用。 43. - kind of process for producing a composition or dosage form used, the process comprising the steps of: (i) the synthetic nanocarriers immunosuppressant conjugated first population; (ii) a second population of the synthetic nano vector with MHC class I-restricted epitopes of an antigen and / or MHC class II restricted epitopes coupling, and (iii) evaluation of these synthetic nanocarriers first and second populations of antigen-specific effector T cells the number of action or activity.
44. 如权利要求43所述的工艺,该工艺包括如在权利要求36-42中任一项所述的方法中所定义的步骤。 44. A process according to claim 43, comprising the process steps of the method as in any one of claims 36-42 as defined.
45. -种通过如权利要求36-44中任一项或如权利要求1-35中任一项所定义的方法或工艺可获得的组合物或剂型。 45. - by species composition or dosage form as claimed method or process as defined in any one of 1 to 35 obtainable according to any one or 36-44 as claimed in claim.
46. -种组合物,包含: (i) 第一群体的合成纳米载体,这些第一群体的合成纳米载体与免疫抑制剂偶联; (ii) 第二群体的合成纳米载体,这些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联;以及(iii) 一种药学上可接受的赋形剂, 其中,该组合物处于在受试者中有效降低抗原特异性效应性T细胞的数目或活性的量。 46. ​​- species composition, comprising: (i) a first population of synthetic nanocarriers, synthetic nanocarriers immunosuppressant coupling these first groups; synthetic nanocarriers (ii) a second population, a second population of these synthetic nanocarriers MHC class I-restricted epitope of an antigen and / or MHC class II restricted epitopes coupled; and a pharmaceutically (iii) one pharmaceutically acceptable excipient, wherein the composition is in amount or activity of the number of antigen-specific effector T cells is effective to reduce the subject.
47. -种供在治疗或预防中使用的组合物,包含: (i) 第一群体的合成纳米载体,这些第一群体的合成纳米载体与免疫抑制剂偶联;以及(ii) 第二群体的合成纳米载体,这些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联; 其中,该组合物处于有效降低在受试者中的抗原特异性效应性T细胞的数目或活性的量。 47. - species for use in the treatment or prevention of a composition comprising: (i) a first population of synthetic nanocarriers, synthetic nanocarriers immunosuppressant coupling these first groups; and (ii) a second population of synthetic nanocarriers, these synthetic nanocarrier second population of MHC class I-restricted epitopes of an antigen and / or MHC II-restricted epitopes coupling; wherein the composition is effective to reduce a subject amount or activity of a number of antigen-specific effector T cells.
48. -种供在一种方法中使用的组合物,包含: (i) 第一群体的合成纳米载体,这些第一群体的合成纳米载体与免疫抑制剂偶联;以及(ii) 第二群体的合成纳米载体,这些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联; 其中,该组合物处于有效降低在受试者中的抗原特异性效应性T细胞的数目或活性的量,该方法是: j .治疗,该治疗包括将所述组合物给予受试者并且在该给予之前和/或之后评定该受试者中的抗原特异性效应性T细胞的数目或活性的步骤; k. 治疗,该治疗包括根据一种治疗方案向受试者给予所述组合物的步骤,该治疗方案先前已经显示降低一个或多个测试受试者中的抗原特异性效应性T细胞的数目或活性; l. 如权利要求1-35中任一项所定义的治疗或预防; m. 例如通过抗原特异性效应性T细胞的数目或活 48. - A method of species for use in the composition, comprising: (i) a first population of synthetic nanocarriers, synthetic nanocarriers immunosuppressant coupling these first groups; and (ii) a second population of synthetic nanocarriers, these synthetic nanocarrier second population of MHC class I-restricted epitopes of an antigen and / or MHC II-restricted epitopes coupling; wherein the composition is effective to reduce a subject the amount of the active or the number of antigen-specific effector T cells, the method is:. j treatment, which treatment comprises administering to a subject the composition and assessment of the subject prior to the administration and / or after step number or activity of effector antigen-specific T cells;. k treatment, the treatment comprising the step of administering to the subject a composition according to a therapeutic regimen, the treatment regimen has previously been shown to reduce one or more antigen-specific effector T cell number or activity of test subject; L 1-35 as defined in any one of the treatment or prevention of claims;. m for example by antigen-specific effector T cells. The number of live or 的降低,对一种炎性疾病、一种自身免疫性疾病、一种过敏症、器官或组织排斥或移植物抗宿主病的治疗或预防; η.对已经经历或将要经历移植的受试者的治疗; 〇.对已经被给予或将要被给予一种治疗性蛋白的受试者的治疗;或Ρ.与一种可移植的移植物或治疗性蛋白相结合的治疗。 Reduction of an inflammatory disease, an autoimmune disease, an allergic disease, organ or tissue rejection or graft versus host disease treatment or prevention; [eta] of the subject has undergone or will undergo transplantation. treatment; square treating the subject has been given or will be given a therapeutic protein;. Ρ with a portable or graft therapeutic protein or therapeutic combination.
49. 一种组合物用于制造供在如权利要求48所述的方法中使用的一种药剂的用途,该组合物包含: (i) 第一群体的合成纳米载体,这些第一群体的合成纳米载体与免疫抑制剂偶联;以及(ii) 第二群体的合成纳米载体,这些第二群体的合成纳米载体与一种抗原的MHC I类限制性表位和/或MHC II类限制性表位偶联; 其中,该组合物处于有效降低在受试者中的抗原特异性效应性T细胞的数目或活性的量。 49. A composition for producing a medicament for use in a method as claimed in claim 48 for use, the composition comprising: (i) a first population of synthetic nanocarriers, the first synthesis of these groups nanocarriers immunosuppressant coupling; synthetic nanocarriers and (ii) a second population, a second population of synthetic nanocarriers of these MHC class I-restricted epitopes of an antigen and / or MHC II-restricted table coupling position; wherein the composition is in an amount effective to reduce the number or activity in the subject antigen-specific effector T cells.
50. 如权利要求48所述的组合物或如权利要求49所述的用途,其中: (a) 该组合物是如权利要求1-35中任一项所定义的;和/或(b) 该组合物是供在一种治疗或预防的方法中使用的,该方法包括通过静脉内、腹膜内、透粘膜、经口、皮下、肺部、鼻内、皮内或肌肉内给予来进行给药,例如,如权利要求12或13中所定义的。 50. The composition of claim 48 or the use as claimed in claim in claim 49, wherein: (a) the composition is any one of 1 to 35 as defined in claim 1; and / or (b) the composition is for use in a method of treatment or prevention of use, the method comprises intravenous, intraperitoneal, transmucosal, oral, subcutaneous, pulmonary, intranasal, intradermal or intramuscular administration to give agents, for example, as set forth in claim 12 or 13, as defined in.
51. -种包含如权利要求45-48和50中任一项所述的组合物的剂型。 51. - kind of dosage form comprising a composition of 45-48 and 50 as claimed in any of claims.
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